Method of treating inflammatory diseases

ABSTRACT

Disclosed are methods of treating inflammatory diseases and disorders, such as arthritis and inflammatory bowel disease, in mammals.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a Continuation-in-Part application of U.S. patentapplication Ser. No. 10/929,295, filed Aug. 30, 2004 now U.S. Pat. No.7,230,099, which is a Continuation-in-Part application of U.S. patentapplication Ser. No. 10/654,580, filed Sep. 3, 2003 now U.S. Pat. No.7,144,907, each of which is incorporated herein by reference in itsentirety.

BACKGROUND OF THE INVENTION

1. Field of the Invention

This invention relates to methods for treating inflammatory diseases anddisorders in mammals, especially humans, which method comprisesadministering an effective amount of a MEK inhibitor to a mammal in needof such treatment.

2. Description of the State of the Art

Cell signaling through growth factor receptors and protein kinases is animportant regulator of cell growth, proliferation and differentiation.In normal cell growth, growth factors, through receptor activation (i.e.PDGF or EGF and others), activate MAP kinase pathways. One of the mostimportant and most well understood MAP kinase pathways involved innormal and uncontrolled cell growth is the Ras/Raf kinase pathway.Active GTP-bound Ras results in the activation and indirectphosphorylation of Raf kinase. Raf then phosphorylates MEK1 and 2 on twoserine residues (S218 and S222 for MEK1 and S222 and S226 for MEK2) (Ahnet al., Methods in Enzymology, 2001, 332, 417-431). Activated MEK thenphosphorylates its only known substrates, the MAP kinases ERK1 and 2.ERK phosphorylation by MEK occurs on Y204 and T202 for ERK1 and Y185 andT183 for ERK2 (Ahn et al., Methods in Enzymology, 2001, 332, 417-431).Phosphorylated ERK dimerizes and then translocates to the nucleus whereit accumulates (Khokhlatchev et al., Cell, 1998, 93, 605-615). In thenucleus, ERK is involved in several important cellular functions,including but not limited to nuclear transport, signal transduction, DNArepair, nucleosome assembly and translocation, and mRNA processing andtranslation (Ahn et al., Molecular Cell, 2000, 6, 1343-1354). Overall,treatment of cells with growth factors leads to the activation of ERK1and 2 which results in proliferation and, in some cases, differentiation(Lewis et al., Adv. Cancer Res., 1998, 74, 49-139).

In proliferative diseases, genetic mutations and/or overexpression ofgrowth factor receptors, downstream signaling proteins, or proteinkinases involved in the ERK kinase pathway lead to uncontrolled cellproliferation and, eventually, tumor formation. For example, somecancers contain mutations which result in the continuous activation ofthis pathway due to continuous production of growth factors. Othermutations can lead to defects in the deactivation of the activatedGTP-bound Ras complex, again resulting in activation of the MAP kinasepathway. Mutated, oncogenic forms of Ras are found in 50% of colonand >90% pancreatic cancers as well as many others types of cancers(Kohl et al., Science, 1993, 260, 1834-1837). Recently, bRaf mutationshave been identified in more than 60% of malignant melanoma (Davies, H.et al., Nature, 2002, 417, 949-954). These mutations in bRaf result in aconstitutively active MAP kinase cascade. Studies of primary tumorsamples and cell lines have also shown constitutive or overactivation ofthe MAP kinase pathway in cancers of pancreas, colon, lung, ovary andkidney (Hoshino, R. et al., Oncogene, 1999, 18, 813-822). Hence, thereis a strong correlation between cancers and an overactive MAP kinasepathway resulting from genetic mutations.

Since constitutive or overactivation of MAP kinase cascade plays apivotal role in cell proliferation and differentiation, inhibition ofthis pathway is believed to be beneficial in hyperproliferativediseases. MEK is a key player in this pathway as it is downstream of Rasand Raf. Additionally, it is an attractive therapeutic target becausethe only known substrates for MEK phosphorylation are the MAP kinases,ERK1 and 2. Inhibition of MEK has been shown to have potentialtherapeutic benefit in several studies. For example, small molecule MEKinhibitors have been shown to inhibit human tumor growth in nude mousexenografts, (Sebolt-Leopold et al., Nature-Medicine, 1999, 5 (7),810-816; Trachet et al., AACR Apr. 6-10, 2002, Poster #5426; Tecle, H.,IBC 2^(nd) International Conference of Protein Kinases, Sep. 9-10,2002), block static allodynia in animals (WO 01/05390 published Jan. 25,2001) and inhibit growth of acute myeloid leukemia cells (Milella etal., J. Clin. Invest., 2001, 108 (6), 851-859).

Small molecule inhibitors of MEK have been recently disclosed. See, forexample, U.S. Pat. No. 5,525,625; WO 98/43960; WO 99/01421; WO 99/01426;WO 00/41505; WO 00/42002; WO 00/42003; WO 00/41994; WO 00/42022; WO00/42029; WO 00/68201; WO 01/68619; and WO 02/06213.

SUMMARY OF THE INVENTION

This invention provides methods for treating an inflammatory disease ordisorder in a mammal, such as a human, which methods compriseadministering a MEK inhibitor to a mammal in need of such treatment.

Examples of MEK inhibitors suitable for use in the methods of thisinvention include compounds of Formula I and solvates, metabolites, andpharmaceutically acceptable salts and prodrugs thereof

wherein:

R¹, R², R⁷, R⁸, R⁹, and R¹⁰ are independently hydrogen, halo, cyano,nitro, trifluoromethyl, difluoromethoxy, trifluoromethoxy, azido, —OR³,—C(O)R³, —C(O)OR³, NR⁴C(O)OR⁶, —OC(O)R³, —NR⁴SO₂R⁶, —SO₂NR³R⁴,—NR⁴C(O)R³, —C(O)NR³R⁴, —NR⁵C(O)NR³R⁴, —NR⁵C(NCN)NR³R⁴, —NR³R⁴, C₁-C₁₀alkyl, C₂-C₁₀ alkenyl, C₂-C₁₀ alkynyl, C₃-C₁₀ cycloalkyl, C₃-C₁₀cycloalkylalkyl, —S(O)_(j)(C₁-C₆ alkyl), —S(O)_(j)(CR⁴R⁵)_(m)-aryl,aryl, arylalkyl, heteroaryl, heteroarylalkyl, heterocyclyl,heterocyclylalkyl, —O(CR⁴R⁵′)_(m)-aryl, —NR⁴(CR⁴R⁵)_(m)-aryl,—O(CR⁴R⁵)_(m)-heteroaryl, —NR⁴(CR⁴R⁵)_(m)-heteroaryl,—O(CR⁴R⁵)_(m)-heterocyclyl or —NR⁴(CR⁴R⁵)_(m)-heterocyclyl, wherein saidalkyl, alkenyl, alkynyl, cycloalkyl, aryl, arylalkyl, heteroaryl,heteroarylalkyl, heterocyclyl and heterocyclylalkyl portions areoptionally substituted with one or more groups independently selectedfrom oxo, halo, cyano, nitro, trifluoromethyl, difluoromethoxy,trifluoromethoxy, azido, —NR⁴SO₂R⁶, —SO₂NR³R⁴, —C(O)R³, —C(O)OR³,—OC(O)R³, —NR⁴C(O)OR⁶, —NR⁴C(O)R³, —C(O)NR³R⁴, —NR³R⁴, —NR⁵C(O)NR³R⁴,—NR⁵C(NCN)NR³R⁴, —OR³, aryl, heteroaryl, arylalkyl, heteroarylalkyl,heterocyclyl, and heterocyclylalkyl;

R³ is hydrogen, trifluoromethyl, C₁-C₁₀ alkyl, C₂-C₁₀ alkenyl, C₂-C₁₀alkynyl, C₃-C₁₀ cycloalkyl, C₃-C₁₀ cycloalkylalkyl, aryl, arylalkyl,heteroaryl, heteroarylalkyl, heterocyclyl, heterocyclylalkyl, phosphate,or an amino acid residue, wherein said alkyl, alkenyl, alkynyl,cycloalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, heterocyclyland heterocyclylalkyl portions are optionally substituted with one ormore groups independently selected from oxo, halo, cyano, nitro,trifluoromethyl, difluoromethoxy, trifluoromethoxy, azido, —NR′SO₂R″″,—SO₂NR′R″, —C(O)R′, C(O)OR′, —OC(O)R′, —NR′C(O)OR″″, —NR′C(O)R″,—C(O)NR′R″, —SR′, —S(O)R″″, —SO₂R″″, —NR′R″, —NR′C(O)NR″R′″,—NR′C(NCN)NR″R′″, —OR′, aryl, heteroaryl, arylalkyl, heteroarylalkyl,heterocyclyl, and heterocyclylalkyl,

or R³ and R⁴ together with the atom to which they are attached form a 4to 10 membered carbocyclic, heteroaryl or heterocyclic ring, whereinsaid carbocyclic, heteroaryl or heterocyclic rings are optionallysubstituted with one or more groups independently selected from halo,cyano, nitro, trifluoromethyl, difluoromethoxy, trifluoromethoxy, azido,—NR′SO₂R″″, —SO₂NR′R″, —C(O)R′, —C(O)OR′, —OC(O)R′, —NR′C(O)OR″″,—NR′C(O)R″, —C(O)NR′R″, —SO₂R″″, —NR′R″, —NR′C(O)NR″R′″,—NR′C(NCN)NR″R′″, —OR′, aryl, heteroaryl, arylalkyl, heteroarylalkyl,heterocyclyl, and heterocyclylalkyl;

R′, R″ and R′″ are independently hydrogen, lower alkyl, lower alkenyl,aryl or arylalkyl, and R″″ is lower alkyl, lower alkenyl, aryl orarylalkyl, or any two of R′, R″, R′″ and R″″ together with the atoms towhich they are attached form a 4 to 10 membered carbocyclic, heteroarylor heterocyclic ring, wherein said alkyl, alkenyl, aryl, arylalkylcarbocyclic rings, heteroaryl rings or heterocyclic rings are optionallysubstituted with one or more groups independently selected from halo,cyano, nitro, trifluoromethyl, difluoromethoxy, trifluoromethoxy, azido,aryl, heteroaryl, arylalkyl, heteroarylalkyl, heterocyclyl, andheterocyclylalkyl;

R⁴ and R⁵ are independently hydrogen or C₁-C₆ alkyl, or

R⁴ and R⁵ together with the atom to which they are attached form a 4 to10 membered carbocyclic, heteroaryl or heterocyclic ring, wherein saidalkyl or said carbocyclic, heteroaryl and heterocyclic rings areoptionally substituted with one or more groups independently selectedfrom halo, cyano, nitro, trifluoromethyl, difluoromethoxy,trifluoromethoxy, azido, —NR′SO₂R′″, —SO₂NR′R″, —C(O)R′″, —C(O)OR′,—OC(O)R′, —NR′C(O)OR″″, —NR′C(O)R″″, —C(O)NR′R″, —SO₂R″″, —NR′R″,—NR″C(O)NR″R′″, —NR′C(NCN)NR″R′″, —OR′, aryl, heteroaryl, arylalkyl,heteroarylalkyl, heterocyclyl and heterocyclylalkyl;

R⁶ is trifluoromethyl, C₁-C₁₀ alkyl, C₃-C₁₀ cycloalkyl, aryl, arylalkyl,heteroaryl, heteroarylalkyl, heterocyclyl, or heterocyclylalkyl, whereinsaid alkyl, cycloalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl,heterocyclyl and heterocyclylalkyl portions are optionally substitutedwith one or more groups independently selected from oxo, halo, cyano,nitro, trifluoromethyl, difluoromethoxy, trifluoromethoxy, azido,—NR′SO₂R″″, —SO₂NR′R″, —C(O)R′, —C(O)OR′, —OC(O)R′, —NR′C(O)OR″″,—NR′C(O)R″, —C(O)NR′R″, —SO₂R″″, —NR′R′, —NR′C(O)NR″R′″,—NR′C(NCN)NR″R′″, —OR′, aryl, heteroaryl, arylalkyl, heteroarylalkyl,heterocyclyl and heterocyclylalkyl;

W is heteroaryl, heterocyclyl, —C(O)OR³, —C(O)NR³R⁴, —C(O)NR⁴OR³,—C(O)R⁴OR³, —C(O)(C₃-C₁₀ cycloalkyl), —C(O)(C₁-C₁₀ alkyl), —C(O)(aryl),—C(O)(heteroaryl), —C(O)(heterocyclyl), —CONH(SO₂)CH₃ or CR³OR³, whereinany of said heteroaryl, heterocyclyl, —C(O)OR³, —C(O)NR³R⁴, —C(O)NR⁴OR³,—C(O)R⁴R³, —C(O)(C₃-C₁₀ cycloalkyl), —C(O)(C₁-C₁₀ alkyl), —C(O)(aryl),—C(O)(heteroaryl), —C(O)(heterocyclyl), —CONH(SO₂)CH₃ and CR³OR³ areoptionally substituted with one or more groups independently selectedfrom —NR³R⁴, —OR³, —R², C, —CIO alkyl, C₂-C₁₀ alkenyl and C₂-C₁₀alkynyl, wherein any of said C₁-C₁₀ alkyl, C₂-C₁₀ alkenyl and C₂-C₁₀alkynyl are optionally substituted with 1 or more groups independentlyselected from —NR³R⁴ and —OR³;

m is 0, 1, 2, 3, 4 or 5;

j is 0, 1 or 2; and

Y is a linker.

Additional examples of MEK inhibitors suitable for use in the methods ofthe present invention include compounds of Formula II and solvates,metabolites, and pharmaceutically acceptable salts and prodrugs thereof

wherein R¹, R², R⁷, R⁸, R⁹, R¹⁰, R″, W, and Y are as defined above, and

R¹¹ is hydrogen, halo, cyano, nitro, trifluoromethyl, difluoromethoxy,trifluoromethoxy, azido, —OR³, —C(O)R³, —C(O)OR³, NR⁴C(O)OR⁶, —OC(O)R³,—NR⁴SO₂R⁶, —SO₂NR³R⁴, —NR⁴C(O)R³, —C(O)NR³R⁴, —NR⁵C(O)NR³R⁴,—NR⁵C(NCN)NR³R⁴, —NR³R⁴, C₁-C₁₀ alkyl, C₂-C₁₀ alkenyl, C₂-C₁₀ alkynyl,C₃-C₁₀ cycloalkyl, C₃-C₁₀ cycloalkylalkyl, —S(O)_(j)(C₁-C₆ alkyl),—S(O)_(j)(CR⁴R⁵)_(m)-aryl, aryl, arylalkyl, heteroaryl, heteroarylalkyl,heterocyclyl, heterocyclylalkyl, —O(CR⁴R⁵)_(m)-aryl,—NR⁴(CR⁴R⁵)_(m)-aryl, —O(CR⁴R⁵)_(m)-heteroaryl,—NR⁴(CR⁴R⁵)_(m)-heteroaryl, —O(CR⁴R⁵)_(m)-heterocyclyl or—NR⁴(CR⁴R⁵)_(m)-heterocyclyl, wherein any of said alkyl, alkenyl,alkynyl, cycloalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl,heterocyclyl and heterocyclylalkyl portions are optionally substitutedwith one or more groups independently selected from oxo, halo, cyano,nitro, trifluoromethyl, difluoromethoxy, trifluoromethoxy, azido,—NR⁴SO₂R⁶, —SO₂NR³R⁴, —C(O)R³, —C(O)OR³, —OC(O)R³, —NR⁴C(O)OR⁶,—NR⁴C(O)R³, —C(O)NR³R⁴, —NR³R⁴, —NR⁵C(O)NR³R⁴, —NR⁵C(NCN)NR³R⁴, —OR³,aryl, heteroaryl, arylalkyl, heteroarylalkyl, heterocyclyl andheterocyclylalkyl

Additional examples of MEK inhibitors suitable for use in the methods ofthe present invention include compounds of Formula III and solvates,metabolites, and pharmaceutically acceptable salts and prodrugs thereof

wherein R¹, R², R⁷, R⁹, R¹⁰, W and Y are as defined above.

Additional examples of MEK inhibitors suitable for use in the methods ofthe present invention include compounds of Formula IV and solvates,metabolites, and pharmaceutically acceptable salts and prodrugs thereof

wherein R¹, R², R⁷, R⁹, R¹⁰, W and Y are as defined above.

Additional examples of MEK inhibitors suitable for use in the methods ofthe present invention include compounds of Formula V and solvates,metabolites, and pharmaceutically acceptable salts and prodrugs thereof

wherein R¹, R², R⁷, R⁸, R⁹, R¹⁰, R¹¹, W and Y are as defined above.

Another aspect of the present invention includes methods for treatinginflammatory disorders in mammals, especially humans, which methodcomprises administering a composition comprising a MEK inhibitor incombination with one or more known therapeutic agents to a mammal inneed of such treatment.

Additional advantages and novel features of this invention shall be setforth in part in the description that follows, and in part will becomeapparent to those skilled in the art upon examination of the followingspecification or may be learned by the practice of the invention. Theadvantages of the invention may be realized and attained by means of theinstrumentalities, combinations, compositions, and methods particularlypointed out in the appended claims.

BRIEF DESCRIPTION OF THE FIGURES

The accompanying drawings, which are incorporated herein and form a partof the specification, illustrate non-limiting embodiments of the presentinvention, and together with the description, serve to explain theprinciples of the invention.

In the Figures:

FIG. 1 is a graph representing mean ankle diameter in inches forestablished type II collagen-induced arthritis in rats with respect totime in days after dosing with AR-14 (1 mg/kg, 3 mg/kg, 10 mg/kg, 30mg/kg or 60 mg/kg), indomethacin (1 mg/kg), ENBREL (etanercept) (10mg/kg), or vehicle.

FIG. 2 is a bar graph representing the mean change in body weight ingrams for established type II collagen-induced arthritis in rats after 7days after dosing with AR-14 (1 mg/kg, 3 mg/kg, 10 mg/kg, 30 mg/kg or 60mg/kg), indomethacin (1 mg/kg), ENBREL (etanercept) 10 mg/kg), orvehicle, relative to normal body weight change.

FIG. 3 is a bar graph representing the mean histopathologic score forankles for established type II collagen-induced arthritis in rats after7 days as a function of the dosage of AR-14 (1 mg/kg, 3 mg/kg, 10 mg/kg,30 mg/kg or 60 mg/kg), indomethacin (1 mg/kg), ENBREL (etanercept) (10mg/kg) or vehicle. Parameters scored were inflammation, pannus,cartilage damage and bone resorption.

FIG. 4 is a graph representing mean ankle diameter in inches foradjuvant-induced arthritis in rats as a function of time in days afterdosing with AR-14 (1 mg/kg, 3 mg/kg, 10 mg/kg, or 30 mg/kg),methotrexate (0.075 mg/kg), or vehicle, relative to normal anklediameter.

FIG. 5 is a bar graph representing mean change in body weight in gramsfor adjuvant-induced arthritis in rats after 15 days as a function ofdosage of AR-14 (1 mg/kg, 3 mg/kg, 10 mg/kg, or 30 mg/kg), methotrexate(0.075 mg/kg), or vehicle, relative to normal body weight.

FIG. 6 is a bar graph representing mean paw weight in grams foradjuvant-induced arthritis in rats after 15 days as a function of thedosage of AR-14 (1 mg/kg, 3 mg/kg, 10 mg/kg, or 30 mg/kg), methotrexate(0.075 mg/kg), or vehicle, relative to normal paw weight.

FIG. 7 is a bar graph representing mean relative spleen weight in gramsfor adjuvant-induced arthritis in rats after 15 days as a function ofthe dosage of AR-14 (1 mg/kg, 3 mg/kg, 10 mg/kg, or 30 mg/kg),methotrexate (0.075 mg/kg), or vehicle, relative to normal spleenweight.

FIG. 8 is a bar graph representing gross lesion score (small intestine)for indomethacin-induced IBD in rats after 4 days as a function of thedosage of AR-14 (1 mg/kg, 3 mg/kg, 10 mg/kg, or 30 mg/kg), dexamethasone(0.1 mg/kg), or vehicle, relative to normal gross lesion score.

FIG. 9 is a bar graph representing mean small intestine weight forindomethacin-induced IBD in rats after 4 days as a function of thedosage of AR-14 (1 mg/kg, 3 mg/kg, 10 mg/kg, or 30 mg/kg), dexamethasone(0.1 mg/kg), or vehicle, relative to normal small intestine weight.

FIG. 10 is a bar graph representing the mean histopathology score forindomethacin-induced IBD in rats after 4 days as a function of thedosage of AR-14 (1 mg/kg, 3 mg/kg, 10 mg/kg, or 30 mg/kg), dexamethasone(0.1 mg/kg), or vehicle, relative to normal histopathology score.Parameters scored were necrosis and inflammation.

FIG. 11 is a bar graph representing mean paw weight in grams in thecarrageenan paw edema model in rats as a function of the dosage of AR-13(3 mg/kg, 10 mg/kg or 30 mg/kg), AR-14 (3 mg/kg, 10 mg/kg, or 30 mg/kg),indomethacin (2 mg/kg), or vehicle, relative to normal paw weight.

FIG. 12 shows the reaction scheme for the synthesis of Compounds 73a and73b.

FIGS. 13A-13G show additional compounds suitable for use in the methodsof the present invention.

DETAILED DESCRIPTION OF THE INVENTION

Disclosed herein are methods for treating an inflammatory disease ordisorder in a mammal, such as a human, which method comprisesadministering a composition including a MEK inhibitor to a mammal inneed of such treatment. In certain embodiments, the methods are used totreat arthritis. In other embodiments, the methods are used to treatinflammatory bowel disease.

As described in detail in the Examples, the effect of a MEK inhibitor ofFormula II, AR-14, was investigated in three animal models of arthritis,namely collagen-induced arthritis (CIA), adjuvant induced arthritis(AIA) (Examples 1 and 2, respectively), and carrageenan paw edema(Example 4). Another MEK inhibitor of Formula II, AR-13, was alsoevaluated in the carrageenan paw edema animal model of inflammation(Example 4).

Oral administration of AR-14 resulted in significant inhibition of theprogression of established arthritis (CIA and AIA models) in rats in adose-dependent manner. In these models, reversal of acute gross andmicroscopic inflammation was observed with once daily oral doses of 10and 30 mg/kg AR-14, respectively. The progression of established diseasewas stopped at 10 mg/kg, and improvement to normal was observed at 60mg/kg in the CIA model and at 30 mg/kg in the AIA model. In addition,significant amelioration of disease (e.g., joint swelling andmicroscopic changes) was seen with once daily oral doses of 10 mg/kgAR-14, and significant inhibition of joint destruction was observed at adose of 1 mg/kg in the CIA model. AR-14 was well tolerated at all dosesadministered and was found to be a selective inhibitor of MEK in vivo,and is useful as a therapeutic tool for arthritis.

In the carrageenan paw edema model, oral administration of AR-13 andAR-14 was observed to significantly inhibit paw weight, and bothcompounds were superior to the positive control indomethacin.

As described in detail in Example 3, a model of Crohn's disease(indomethacin-induced intestinal injury) was utilized to evaluate theeffectiveness of AR-14 in treating inflammatory bowel disease (IBD).Oral administration of AR-14 resulted in significant inhibition of grossand microscopic gut inflammation in the indomethacin-inducedinflammatory bowel disease model in rats, as measured by gut score,weight changes, and microscopic changes, with once daily oral doses of10 mg/kg AR-14. AR-14 was well tolerated at all doses administered andwas found to be a selective inhibitor of MEK in vivo, and is useful as atherapeutic tool for IBD.

Accordingly, one aspect of the invention features the use of MEKinhibitors in the treatment of inflammatory diseases and disorders.Examples of MEK inhibitors suitable for use in the methods of thisinvention include compounds of Formula I and solvates, metabolites, andpharmaceutically acceptable salts and prodrugs thereof

wherein:

R¹, R², R⁷, R⁸, R⁹, and R¹⁰ are independently hydrogen, halo, cyano,nitro, trifluoromethyl, difluoromethoxy, trifluoromethoxy, azido, —OR³,—C(O)R³, —C(O)OR³, NR⁴C(O)OR⁶, —OC(O)R³, —NR⁴SO₂R⁶, —SO₂NR³R⁴,—NR⁴C(O)R³, —C(O)NR³R⁴, —NR⁵C(O)NR³R⁴, —NR⁵C(NCN)NR³R⁴, —NR³R⁴, C₁-C₁₀alkyl, C₂-C₁₀ alkenyl, C₂-C₁₀ alkynyl, C₃-C₁₀ cycloalkyl, C₃-C₁₀cycloalkylalkyl, —S(O)_(j)(C₁-C₆ alkyl), —S(O)_(j)(CR⁴R⁵)_(m)-aryl,aryl, arylalkyl, heteroaryl, heteroarylalkyl, heterocyclyl,heterocyclylalkyl, —O(CR⁴R⁵)_(m)-aryl, —NR⁴(CR⁴R⁵)_(m)-aryl,—O(CR⁴R⁵)_(m)-heteroaryl, —NR⁴(CR⁴R⁵)_(m)-heteroaryl,—O(CR⁴R⁵)_(m)-heterocyclyl or —NR⁴(CR⁴R⁵)_(m)-heterocyclyl, wherein saidalkyl, alkenyl, alkynyl, cycloalkyl, aryl, arylalkyl, heteroaryl,heteroarylalkyl, heterocyclyl and heterocyclylalkyl portions areoptionally substituted with one or more groups independently selectedfrom oxo, halo, cyano, nitro, trifluoromethyl, difluoromethoxy,trifluoromethoxy, azido, —NR⁴SO₂R⁶, —SO₂NR³R⁴, —C(O)R³, —C(O)OR³,—OC(O)R³, —NR⁴C(O)OR⁶, —NR⁴C(O)R³, —C(O)NR³R⁴, —NR³R⁴, —NR⁵C(O)NR³R⁴,—NR⁵C(NCN)NR³R⁴, —OR³, aryl, heteroaryl, arylalkyl, heteroarylalkyl,heterocyclyl and heterocyclylalkyl;

R³ is hydrogen, trifluoromethyl, C₁-C₁₀ alkyl, C₂-C₁₀ alkenyl, C₂-C₁₀alkynyl, C₃-C₁₀ cycloalkyl, C₃-C₁₀ cycloalkylalkyl, aryl, arylalkyl,heteroaryl, heteroarylalkyl, heterocyclyl, heterocyclylalkyl, phosphate,or an amino acid residue, wherein said alkyl, alkenyl, alkynyl,cycloalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, heterocyclyland heterocyclylalkyl portions are optionally substituted with one ormore groups independently selected from oxo, halo, cyano, nitro,trifluoromethyl, difluoromethoxy, trifluoromethoxy, azido, —NR′SO₂R″″,—SO₂NR′R″, —C(O)R′, C(O)OR′, —OC(O)R′, —NR′C(O)OR″″, —NR′C(O)R″,—C(O)NR′R″, —SR′, —S(O)R″″, —SO₂R″″, —NR′R″, —NR′C(O)NR″R′″,—NR′C(NCN)NR″R′″, —OR′, aryl, heteroaryl, arylalkyl, heteroarylalkyl,heterocyclyl and heterocyclylalkyl,

or R³ and R⁴ together with the atom to which they are attached form a 4to 10 membered carbocyclic, heteroaryl or heterocyclic ring, whereinsaid carbocyclic, heteroaryl or heterocyclic rings are optionallysubstituted with one or more groups independently selected from halo,cyano, nitro, trifluoromethyl, difluoromethoxy, trifluoromethoxy, azido,—NR′SO₂R″″, —SO₂NR′R″, —C(O)R′, —C(O)OR′, —OC(O)R′, —NR′C(O)OR″″,—NR′C(O)R″, —C(O)NR′R″, —SO₂R″″, —NR′R″, —NR′C(O)NR″R′″,—NR′C(NCN)NR″R′″, —OR′, aryl, heteroaryl, arylalkyl, heteroarylalkyl,heterocyclyl and heterocyclylalkyl;

R′, R″ and R′″ are independently hydrogen, lower alkyl, lower alkenyl,aryl or arylalkyl, and R″″ is lower alkyl, lower alkenyl, aryl orarylalkyl, or any two of R′, R″, R′″ and R″″ together with the atoms towhich they are attached form a 4 to 10 membered carbocyclic, heteroarylor heterocyclic ring, wherein said alkyl, alkenyl, aryl, arylalkylcarbocyclic rings, heteroaryl rings or heterocyclic rings are optionallysubstituted with one or more groups independently selected from halo,cyano, nitro, trifluoromethyl, difluoromethoxy, trifluoromethoxy, azido,aryl, heteroaryl, arylalkyl, heteroarylalkyl, heterocyclyl, andheterocyclylalkyl;

R⁴ and R⁵ are independently hydrogen or C₁-C₆ alkyl, or

R⁴ and R⁵ together with the atom to which they are attached form a 4 to10 membered carbocyclic, heteroaryl or heterocyclic ring, wherein saidalkyl or said carbocyclic, heteroaryl and heterocyclic rings areoptionally substituted with one or more groups independently selectedfrom halo, cyano, nitro, trifluoromethyl, difluoromethoxy,trifluoromethoxy, azido, —NR′SO₂R″″, —SO₂NR′R″, —C(O)R″″, —C(O)OR′,—OC(O)R′, —NR′C(O)OR″″, —NR′C(O)R″, —C(O)NR′R″, —SO₂R″″, —NR′R″,—NR′C(O)NR″R′″, —NR′C(NCN)NR″R′″, —OR′, aryl, heteroaryl, arylalkyl,heteroarylalkyl, heterocyclyl and heterocyclylalkyl;

R⁶ is trifluoromethyl, C₁-C₁₀ alkyl, C₃-C₁₀ cycloalkyl, aryl, arylalkyl,heteroaryl, heteroarylalkyl, heterocyclyl or heterocyclylalkyl, whereinsaid alkyl, cycloalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl,heterocyclyl and heterocyclylalkyl portions are optionally substitutedwith one or more groups independently selected from oxo, halo, cyano,nitro, trifluoromethyl, difluoromethoxy, trifluoromethoxy, azido,—NR′SO₂R″″, —SO₂NR′R″, —C(O)R′, —C(O)OR′, —OC(O)R′, —NR′C(O)OR″″,—NR′C(O)R″, —C(O)NR′R″, —SO₂R″″, —NR′R′, —NR′C(O)NR″R′″,—NR′C(NCN)NR″R′″, —OR′, aryl, heteroaryl, arylalkyl, heteroarylalkyl,heterocyclyl and heterocyclylalkyl;

W is heteroaryl, heterocyclyl, —C(O)OR³, —C(O)NR³R⁴, —C(O)NR⁴OR³,—C(O)R⁴OR³, —C(O)(C₃-C₁₀ cycloalkyl), —C(O)(C₁-C₁₀ alkyl), —C(O)(aryl),—C(O)(heteroaryl), —C(O)(heterocyclyl), —CONH(SO₂)CH₃ or CR³OR³, whereinany of said heteroaryl, heterocyclyl, —C(O)OR³, —C(O)NR³R⁴, —C(O)NR⁴OR³,—C(O)R⁴OR³, —C(O)(C₃-C₁₀ cycloalkyl), —C(O)(C₁-C₁₀ alkyl), —C(O)(aryl),—C(O)(heteroaryl), —C(O)(heterocyclyl), —CONH(SO₂)CH₃ and CR³OR³ areoptionally substituted with one or more groups independently selectedfrom —NR³R⁴, —OR³, —R², C₁-C₁₀ alkyl, C₂-C₁₀ alkenyl and C₂-C₁₀ alkynyl,wherein any of said C₁-C₁₀ alkyl, C₂-C₁₀ alkenyl, and C₂-C₁₀ alkynyl areoptionally substituted with 1 or more groups independently selected from—NR³R⁴ and —OR³;

m is 0, 1, 2, 3, 4 or 5;

j is 0, 1 or 2; and

Y is a linker.

A “linker” is a molecular entity that connects two or more molecularentities through covalent or non-covalent interactions. Examples oflinkers include, but are not limited to, NR³, O, S, S(O), S(O)₂, C(O),and CH₂, where R³ is as defined above. In certain embodiments, themethod utilizes a compound of Formula I wherein Y is NR³. In particularembodiments, Y is NH. In certain embodiments, W is —C(O)OR³,—C(O)NR⁴OR³, or —CONH(SO₂)CH₃.

Additional examples of MEK inhibitors suitable for use in the methods ofthe present invention include compounds of Formula II solvates,metabolites, and pharmaceutically acceptable salts and prodrugs thereof

wherein R¹, R², R⁷, R⁸, R⁹, R¹⁰, R¹¹, W and Y are as defined above, and

R¹¹ is hydrogen, halo, cyano, nitro, trifluoromethyl, difluoromethoxy,trifluoromethoxy, azido, —OR³, —C(O)R³, —C(O)OR³, NR⁴C(O)OR⁶, —OC(O)R³,—NR⁴SO₂R⁶, —SO₂NR³R⁴⁴—NR⁴C(O)R³, —C(O)NR³R⁴, —NR⁵C(O)NR³R⁴,—NR⁵C(CN)NR³R⁴, —NR³R⁴, C₁-C₁₀ alkyl, C₂-C₁₀ alkenyl, C₂-C₁₀ alkynyl,C₃-C₁₀ cycloalkyl, C₃-C₁₀ cycloalkylalkyl, —S(O)_(j)(C₁-C₆ alkyl),—S(O)_(j)(CR⁴R⁵)_(m)-aryl, aryl, arylalkyl, heteroaryl, heteroarylalkyl,heterocyclyl, heterocyclylalkyl, —O(CR⁴R⁵)_(m)-aryl,—NR⁴(CR⁴R⁵)_(m)-aryl, —O(CR⁴R⁵)_(m)-heteroaryl,—NR⁴(CR⁴R⁵)_(m)-heteroaryl, —O(CR⁴R⁵)_(m)-heterocyclyl or—NR⁴(CR⁴R⁵)_(m)-heterocyclyl, wherein any of said alkyl, alkenyl,alkynyl, cycloalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl,heterocyclyl and heterocyclylalkyl portions are optionally substitutedwith one or more groups independently selected from oxo, halo, cyano,nitro, trifluoromethyl, difluoromethoxy, trifluoromethoxy, azido,—NR⁴SO₂R⁶, —SO₂NR³R⁴, —C(O)R³, —C(O)OR³, —OC(O)R³, —NR⁴C(O)OR⁶,—NR⁴C(O)R³, —C(O)NR³R⁴, —NR³R⁴, —NR⁵C(O)NR³R⁴, —NR⁵C(NCN)NR³R⁴, —OR³,aryl, heteroaryl, arylalkyl, heteroarylalkyl, heterocyclyl andheterocyclylalkyl. In certain embodiments, the method utilizes acompound of Formula II wherein Y is NR³. In particular embodiments, Y isNH. In certain embodiments, W is —C(O)OR³, —C(O)NR⁴OR³, or—CONH(SO₂)CH₃.

Additional examples of MEK inhibitors suitable for use in the methods ofthe present invention include compounds of Formula III and solvates,metabolites, and pharmaceutically acceptable salts and prodrugs thereof

wherein R¹, R², R⁷, R⁸, R⁹, R¹⁰, W and Y are as defined above. Incertain embodiments, the method utilizes a compound of Formula IIIwherein Y is NR³. In particular embodiments, Y is NH. In certainembodiments, W is —C(O)OR³, —C(O)NR⁴OR³, or —CONH(SO₂)CH₃.

Additional examples of MEK inhibitors suitable for use in the methods ofthe present invention include compounds of Formula IV and solvates,metabolites, and pharmaceutically acceptable salts and prodrugs thereof

wherein R¹, R², R⁷, R⁸, R⁹, R¹⁰, W and Y are as defined above. Incertain embodiments, the method utilizes a compound of Formula IVwherein Y is NR³. In particular embodiments, Y is NH. In certainembodiments, W is —C(O)OR³, —C(O)NR⁴OR³, or —CONH(SO₂)CH₃.

Additional examples of MEK inhibitors suitable for use in the methods ofthe present invention include compounds of Formula V and solvates,metabolites, and pharmaceutically acceptable salts and prodrugs thereof

wherein R¹, R², R⁷, R⁸, R⁹, R¹⁰, R¹¹, W and Y are as defined above. Incertain embodiments, the method utilizes a compound of Formula V whereinY is NR³. In particular embodiments, Y is NH. In certain embodiments, Wis —C(O)OR³, —C(O)NR⁴OR³, or —CONH(SO₂)CH₃.

The terms “alkyl” and “lower alkyl” as used herein refer to a saturatedlinear or branched-chain monovalent hydrocarbon radical having one toten carbon atoms, wherein the alkyl radical may be optionallysubstituted independently with one or more substituents described below.Examples of alkyl groups include, but are not limited to, methyl, ethyl,n-propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl,isopentyl, neopentyl, tert-pentyl, hexyl, 2-hexyl, 3-hexyl,3-methylpentyl, heptyl, octyl, and the like.

The terms “alkenyl”, “lower alkenyl” and “alkenyl” refer to linear orbranched-chain monovalent hydrocarbon radical having two to 10 carbonatoms and at least one double bond, and include, but is not limited to,ethenyl, propenyl, 1-but-3-enyl, 1-pent-3-enyl, 1-hex-5-enyl and thelike, wherein the alkenyl radical may be optionally substitutedindependently with one or more substituents described herein, andincludes radicals having “cis” and “trans” orientations, oralternatively, “E” and “Z” orientations.

The terms “lower alkynyl” and “alkynyl” refer to a linear or branchedmonovalent hydrocarbon radical of two to twelve carbon atoms containingat least one triple bond. Examples include, but are not limited to,ethynyl, propynyl, butynyl, pentyn-2-yl and the like, wherein thealkynyl radical may be optionally substituted independently with one ormore substituents described herein.

The term “allyl” refers to a radical having the formula RC═CHCHR,wherein R is alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl,aryl, heteroaryl, or any substituent as defined herein, wherein theallyl may be optionally substituted independently with one or moresubstituents described herein.

The terms “carbocycle,” “carbocyclyl,” and “cycloalkyl” refer tosaturated or partially unsaturated cyclic hydrocarbon radical havingfrom three to ten carbon atoms. The term “cycloalkyl” includesmonocyclic and polycyclic (e.g., bicyclic and tricyclic) cycloalkylstructures, wherein the polycyclic structures optionally include asaturated or partially unsaturated cycloalkyl fused to a saturated orpartially unsaturated cycloalkyl or heterocycloalkyl ring or an aryl orheteroaryl ring. Examples of cycloalkyl groups include, but are notlimited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,cycloheptyl, and the like. The cycloalkyl may be optionally substitutedindependently in one or more substitutable positions with variousgroups. For example, such cycloalkyl groups may be optionallysubstituted with, for example, C₁-C₆ alkyl, C₁-C₆ alkoxy, halo, hydroxy,cyano, nitro, amino, mono(C₁-C₆)alkylamino, di(C₁-C₆)alkylamino, C₂-C₆alkenyl, C₂-C₆ alkynyl, C₁-C₆ haloalkyl, C₁-C₆ haloalkoxy,amino(C₁-C₆)alkyl, mono(C₁-C₆)alkylamino(C₁-C₆)alkyl ordi(C₁-C₆)alkylamino(C₁-C₆)alkyl.

The term “heteroalkyl” refers to saturated linear or branched-chainmonovalent hydrocarbon radical of one to twelve carbon atoms, wherein atleast one of the carbon atoms is replaced with a heteroatom selectedfrom N, O, or S, and wherein the radical may be a carbon radical orheteroatom radical (i.e., the heteroatom may appear in the middle or atthe end of the radical). The heteroalkyl radical may be optionallysubstituted independently with one or more substituents describedherein. The term “heteroalkyl” encompasses alkoxy and heteroalkoxyradicals.

The terms “heterocycloalkyl,” “heterocycle” or “hetercyclyl” refer to asaturated or partially unsaturated carbocyclic radical of 3 to 8 ringatoms in which at least one ring atom is a heteroatom selected fromnitrogen, oxygen and sulfur, the remaining ring atoms being C, whereinone or more ring atoms may be optionally substituted independently withone or more substituent described below. The radical may be a carbonradical or heteroatom radical. The term further includes fused ringsystems which include a heterocycle fused to an aromatic group.“Heterocycloalkyl” also includes radicals where heterocycle radicals arefused with aromatic or heteroaromatic rings. Examples ofheterocycloalkyl rings include, but are not limited to, pyrrolidinyl,tetrahydrofuranyl, dihydrofuranyl, tetrahydrothienyl, tetrahydropyranyl,dihydropyranyl, tetrahydrothiopyranyl, piperidino, morpholino,thiomorpholino, thioxanyl, piperazinyl, homopiperazinyl, azetidinyl,oxetanyl, thietanyl, homopiperidinyl, oxepanyl, thiepanyl, oxazepinyl,diazepinyl, thiazepinyl, 1,2,3,6-tetrahydropyridinyl, 2-pyrrolinyl,3-pyrrolinyl, indolinyl, 2H-pyranyl, 4H-pyranyl, dioxanyl,1,3-dioxolanyl, pyrazolinyl, dithianyl, dithiolanyl, dihydropyranyl,dihydrothienyl, dihydrofuranyl, pyrazolidinylimidazolinyl,imidazolidinyl, 3-azabicyco[3.1.0]hexanyl, 3-azabicyclo[4.1.0]heptanyl,azabicyclo[2.2.2]hexanyl, 3H-indolyl and quinolizinyl. Spiro moietiesare also included within the scope of this definition. The foregoinggroups, as derived from the groups listed above, may be C-attached orN-attached where such is possible. For instance, a group derived frompyrrole may be pyrrol-1-yl (N-attached) or pyrrol-3-yl (C-attached).Further, a group derived from imidazole may be imidazol-1-yl(N-attached) or imidazol-3-yl (C-attached). An example of a heterocyclicgroup wherein 2 ring carbon atoms are substituted with oxo (═O) moietiesis 1,1-dioxo-thiomorpholinyl. The heterocycle groups herein areunsubstituted or, as specified, substituted in one or more substitutablepositions with various groups. For example, such heterocycle groups maybe optionally substituted with, for example, C₁-C₆ alkyl, C₁-C₆ alkoxy,halo, hydroxy, cyano, nitro, amino, mono(C₁-C₆)alkylamino,di(C₁-C₆)alkylamino, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₁-C₆ haloalkyl,C₁-C₆ haloalkoxy, amino(C₁-C₆)alkyl, mono(C₁-C₆)alkylamino(C₁-C₆)alkylor di(C₁-C₆)alkylamino(C₁-C₆)alkyl.

The term “aryl” refers to a monovalent aromatic carbocyclic radicalhaving a single ring (e.g., phenyl), multiple rings (e.g., biphenyl), ormultiple condensed rings in which at least one is aromatic, (e.g.,1,2,3,4-tetrahydronaphthyl, naphthyl), which is optionally mono-, di-,or trisubstituted with, e.g., halo, lower alkyl, lower alkoxy,trifluoromethyl, aryl, heteroaryl, and hydroxy.

The term “heteroaryl” refers to a monovalent aromatic radical of 5-, 6-,or 7-membered rings which includes fused ring systems (at least one ofwhich is aromatic) of 5-10 atoms containing at least one and up to fourheteroatoms selected from nitrogen, oxygen, or sulfur. Examples ofheteroaryl groups are pyridinyl, imidazolyl, pyrimidinyl, pyrazolyl,triazolyl, pyrazinyl, tetrazolyl, furyl, thienyl, isoxazolyl, thiazolyl,oxazolyl, isothiazolyl, pyrrolyl, quinolinyl, isoquinolinyl, indolyl,benzimidazolyl, benzofuranyl, cinnolinyl, indazolyl, indolizinyl,phthalazinyl, pyridazinyl, triazinyl, isoindolyl, pteridinyl, purinyl,oxadiazolyl, triazolyl, thiadiazolyl, thiadiazolyl, furazanyl,benzofurazanyl, benzothiophenyl, benzothiazolyl, benzoxazolyl,quinazolinyl, quinoxalinyl, naphthyridinyl, and furopyridinyl. Spiromoieties are also included within the scope of this definition.Heteroaryl groups are optionally mono-, di-, or trisubstituted with,e.g., halo, lower alkyl, lower alkoxy, haloalkyl, aryl, heteroaryl, andhydroxy.

The term “halo” includes fluoro, bromo, chloro, and iodo substituents.

The term “arylalkyl” means an alkyl moiety (as defined above)substituted with one or more aryl moiety (also as defined above).Examples of arylalkyl radicals are aryl-C₁₋₃-alkyls including, but notlimited to, benzyl, phenylethyl, and the like.

The term “heteroarylalkyl” means an alkyl moiety (as defined above)substituted with a heteroaryl moiety (also as defined above). Examplesinclude 5- or 6-membered heteroaryl-C₁₋₃-alkyls such as, but not limitedto, oxazolylmethyl, pyridylethyl and the like.

The term “heterocyclylalkyl” means an alkyl moiety (as defined above)substituted with a heterocyclyl moiety (also defined above). Examplesinclude 5- or 6-membered heterocyclyl-C₁₋₃-alkyls such as, but notlimited to, tetrahydropyranylmethyl.

The term “cycloalkylalkyl” means an alkyl moiety (as defined above)substituted with a cycloalkyl moiety (also defined above). Examplesinclude 5- or 6membered cycloalkyl-C₁₋₃-alkyls such ascyclopropylmethyl.

The term “Me” means methyl, “Et” means ethyl, “Bu” means butyl and “Ac”means acetyl.

In general, the various moieties or functional groups of the compoundsof Formulas I-V may be optionally substituted by one or moresubstituents. Examples of substituents suitable for purposes of thisinvention include, but are not limited to, oxo, halo, cyano, nitro,trifluoromethyl, difluoromethoxy, trifluoromethoxy, azido, —NR⁴SO₂R⁶,—SO₂NR³R⁴, —C(O)R³, —C(O)OR³, —OC(O)R³, —NR⁴C(O)OR⁶, —NR⁴C(O)R³,—C(O)NR³R⁴, —NR³R⁴, —NR⁵C(O)NR³R⁴, —NR⁵C(NCN)NR³R⁴, —OR³, aryl,heteroaryl, arylalkyl, heteroarylalkyl, heterocyclyl, andheterocyclylalkyl, wherein R³, R⁴, R⁵ and R⁶ are as defined herein.

It is to be understood that in instances where two or more radicals areused in succession to define a substituent attached to a structure, thefirst named radical is considered to be terminal and the last namedradical is considered to be attached to the structure in question. Thus,for example, the radical arylalkyl is attached to the structure inquestion by the alkyl group.

In the above compounds, where a term such as (CR⁴R⁵)_(m) is used, R⁴ andR⁵ may vary with each iteration of m above 1. For instance, where m is2, the term (CR⁴R⁵)_(m) may equal —CH₂CH₂— or—CH(CH₃)C(CH₂CH₃)(CH₂CH₂CH₃)— or any number of similar moieties fallingwithin the scope of the definitions of R⁴ and R⁵.

The compounds of Formulas I-V may possess one or more asymmetriccenters; such compounds can therefore be produced as individual (R)- or(S)-stereoisomers or as mixtures thereof. Unless indicated otherwise,the description or naming of a particular compound in the specificationand claims is intended to include both individual enantiomers,diastereomers mixtures, racemic or otherwise, thereof. Accordingly, thisinvention also includes the use of such isomers, includingdiastereomeric mixtures and pure enantiomers of the compounds ofFormulas I-V, in the treatment of inflammatory disorders and diseases.

This invention also encompasses the use of pharmaceutically acceptableprodrugs of compounds of Formulas I-V in the treatment of inflammatorydisorders. A “pharmaceutically acceptable prodrug” is a compound thatmay be converted under physiological conditions or by solvolysis to thespecified compound or to a pharmaceutically acceptable salt of suchcompound. Prodrugs include compounds wherein an amino acid residue, or apolypeptide chain of two or more (e.g., two, three or four) amino acidresidues is covalently joined through an amide or ester bond to a freeamino, hydroxy or carboxylic acid group of compounds of Formulas I-V.The amino acid residues include but are not limited to the 20 naturallyoccurring amino acids commonly designated by three letter symbols andalso includes 4-hydroxyproline, hydroxylysine, demosine, isodemosine,3-methylhistidine, norvaline, beta-alanine, gamma-aminobutyric acid,cirtulline, homocysteine, homoserine, ornithine and methionine sulfone.One embodiment of a prodrug suitable for use in the methods of thepresent invention is a compound of Formula I-V covalently joined to aphosphate residue. Another example of a suitable prodrug is a compoundof Formula I-V covalently joined to a valine residue.

Additional types of prodrugs are also encompassed. For instance, freecarboxyl groups can be derivatized as amides or alkyl esters. Freehydroxy groups may be derivatized using groups including but not limitedto phosphate esters, hemisuccinates, dimethylaminoacetates, andphosphoryloxymethyloxycarbonyls, as outlined in Advanced Drug DeliveryReviews, 1996, 19, 115. Carbamate prodrugs of hydroxy and amino groupsare also included, as are carbonate prodrugs, sulfonate esters andsulfate esters of hydroxy groups. Derivatization of hydroxy groups as(acyloxy)methyl and (acyloxy)ethyl ethers wherein the acyl group may bean alkyl ester, optionally substituted with groups including but notlimited to ether, amine and carboxylic acid functionalities, or whereinthe acyl group is an amino acid ester as described above, are alsoencompassed. Prodrugs of this type are described in J. Med. Chem., 1996,39, 10. Free amines can also be derivatized as amides, sulfonamides orphosphonamides. All of these prodrug moieties may incorporate groupsincluding but not limited to ether, amine and carboxylic acidfunctionalities.

In addition, the invention also includes the use of solvates,pharmaceutically active metabolites, and pharmaceutically acceptablesalts of compounds of Formulas I-V.

The term “solvate” refers to an aggregate of a compound of Formula I-Vwith one or more solvent molecules.

A “metabolite” is a pharmacologically active product produced through invivo metabolism in the body of a compound of Formula I-V or a saltthereof. Such products may result for example from the oxidation,reduction, hydrolysis, amidation, deamidation, esterification,deesterification, enzymatic cleavage, and the like, of the administeredcompound. Metabolites are typically identified by preparing aradiolabelled (e.g., ¹⁴C or ³H) isotope of a compound of the invention,administering it parenterally in a detectable dose (e.g., greater thanabout 0.5 mg/kg) to an animal such as rat, mouse, guinea pig, monkey, orto man, allowing sufficient time for metabolism to occur (typicallyabout 30 seconds to about 30 hours) and isolating its conversionproducts from the urine, blood or other biological samples. Theseproducts are easily isolated since they are labeled (others are isolatedby the use of antibodies capable of binding epitopes surviving in themetabolite). The metabolite structures are determined in conventionalfashion, e.g., by MS, LC/MS or NMR analysis. In general, analysis ofmetabolites is done in the same way as conventional drug metabolismstudies well known to those skilled in the art. The metabolites, so longas they are not otherwise found in vivo, are useful in diagnostic assaysfor therapeutic dosing of the compounds of the invention.

A “pharmaceutically acceptable salt” as used herein, unless otherwiseindicated, includes salts that retain the biological effectiveness ofthe free acids and bases of the specified compound and that are notbiologically or otherwise undesirable. A compound of Formula I-V maypossess a sufficiently acidic, a sufficiently basic, or both functionalgroups, and accordingly react with any of a number of inorganic ororganic bases, and inorganic and organic acids, to form apharmaceutically acceptable salt. Examples of pharmaceuticallyacceptable salts include those salts prepared by reaction of thecompounds of the present invention with a mineral or organic acid or aninorganic base, such salts including sulfates, pyrosulfates, bisulfates,sulfites, bisulfites, phosphates, monohydrogenphosphates,dihydrogenphosphates, metaphosphates, pyrophosphates, chlorides,bromides, iodides, acetates, propionates, decanoates, caprylates,acrylates, formates, isobutyrates, caproates, heptanoates, propiolates,oxalates, malonates, succinates, suberates, sebacates, fumarates,maleates, butyn-1,4-dioates, hexyne-1,6-dioates, benzoates,chlorobenzoates, methylbenzoates, dinitrobenzoates, hydroxybenzoates,methoxybenzoates, phthalates, sulfonates, xylenesulfonates,phenylacetates, phenylpropionates, phenylbutyrates, citrates, lactates,γ-hydroxybutyrates, glycollates, tartrates, methanesulfonates,propanesulfonates, naphthalene-1-sulfonates, naphthalene-2-sulfonates,and mandelates. Since a single compound of the present invention mayinclude more than one acidic or basic moiety, the compounds of thepresent invention may include mono, di or tri-salts in a singlecompound.

If the compound of Formula I-V is a base, the desired pharmaceuticallyacceptable salt may be prepared by any suitable method available in theart, for example, treatment of the free base with an acidic compound,particularly an inorganic acid, such as hydrochloric acid, hydrobromicacid, sulfuric acid, nitric acid, phosphoric acid and the like, or withan organic acid, such as acetic acid, maleic acid, succinic acid,mandelic acid, fumaric acid, malonic acid, pyruvic acid, oxalic acid,glycolic acid, salicylic acid, a pyranosidyl acid, such as glucuronicacid or galacturonic acid, an alpha-hydroxy acid, such as citric acid ortartaric acid, an amino acid, such as aspartic acid or glutamic acid, anaromatic acid, such as benzoic acid or cinnamic acid, a sulfonic acid,such as p-toluenesulfonic acid or ethanesulfonic acid, or the like.

If the compound of Formula I-V is an acid, the desired pharmaceuticallyacceptable salt may be prepared by any suitable method, for example,treatment of the free acid with an inorganic or organic base. In certainembodiments, inorganic salts are those formed with alkali and alkalineearth metals such as lithium, sodium, potassium, barium and calcium.Examples of organic base salts include, for example, ammonium,dibenzylammonium, benzylammonium, 2-hydroxyethylammonium,bis(2-hydroxyethyl)ammonium, phenylethylbenzylamine,dibenzylethylenediamine, and the like salts. Other salts of acidicmoieties may include, for example, those salts formed with procaine,quinine and N-methylglusoamine, plus salts formed with basic amino acidssuch as glycine, ornithine, histidine, phenylglycine, lysine andarginine.

The compounds of Formula I-V may be prepared using the reaction routesand synthesis schemes 1-9 as described below, employing the techniquesavailable in the art using starting materials that are readily availableor can be synthesized using methods known in the art.

Scheme 1 illustrates a method of preparing compounds of the Formula I.Carboxylic acid 102 can be prepared from arene 101 by deprotonation atlow temperature (−100 to −60° C.) in the appropriate ethereal solventsuch as THF or diethyl ether followed by carbon dioxide quench, whichcan be performed with solid dry ice. The deprotonation can beaccomplished with LDA in THF at −78° C. In one example, the quenchmethod comprises adding the aryllithium THF solution via cannula to asaturated solution of dry carbon dioxide in THF at −78° C. and thenwarming to room temperature. Aniline 103 can be prepared bydeprotonation of an appropriate 2-substituted aniline with KHMDS,LiHMDS, NaHMDS or LDA at low temperature (−100 to −60° C.) in anappropriate ethereal solvent such as THF or diethyl ether, followed byaddition of carboxylic acid 102 and warming to room temperature. In oneembodiment, deprotonation is accomplished with LDA at −78° C. in THF,followed by addition of carboxylic acid 102 and warming to roomtemperature. Ester 104 can be prepared by standard methods including,but not limited to, Fisher esterification (MeOH, H₂SO₄), reaction withTMSCHN₂ or TMSCl in MeOH. Acetylene derivative 105 is prepared bySonagashria coupling of bromide 104 using an appropriately substitutedacetylene, CuI, an amine base, palladium catalyst and organic solventsuch as DME, THF, or DMF at temperatures between 25 and 100° C. Suitablepalladium catalysts include, but are not limited to, PdCl₂(dppf),Pd(Ph₃P)₄, and Pd₂ dba₃/dppf. Suitable amine bases include, but are notlimited, to Et₃N, Hunig's base, and diisopropylamine. In one embodiment,the Pd(0) mediated coupling to prepare acetylene 105 is accomplishedwith Pd(PPh₃)₂Cl₂, CuI, diisopropylamine, and the appropriatesubstituted acetylene in THF at room temperature. Hydrolysis ofacetylene 105 to prepare ketone 106 can be accomplished by standardmethods including but not limited to H₂SO₄, TFA, trifluorosulfonamide,FeCl₃, or HgSO₄/H₂SO₄. Benzisoxazole 107 can be prepared in a two-stepprocedure from ketone 106. Addition of the potassium salt of acetoneoxime in suitable organic solvent such as THF or Et₂O at temperaturesranging from −78 to 5° C. is followed by acid catalyzed cyclization. Theacetone oxime addition is most easily performed by addition of a THFsolution of ketone 106 to the salt at 0° C. The cyclization can beaccomplished using a variety of acidic aqueous conditions at a range oftemperatures. In one embodiment cyclization is accomplished by treatmentof the isopropylideneaminooxybenzoic acid methyl ester with 5% aqueousHCl in MeOH at reflux. Halogenation to form benzisoxazole 108 isaccomplished using standard procedures such as NCS or NBS in DMF.Hydrolysis of ester 108 to form carboxylic acid 109 can be performedunder standard conditions. The acid can be converted to hydroxamate 110or amide 112 by standard coupling procedures including but not limitedto EDCI/HOBt, PyBOP, or DIC and the appropriate hydroxylamine or amine.Alternatively, hydroxamate 110 or amide 112 can be prepared in two stepsby initial conversion to the acid chloride by standard methods followedby addition of the hydroxylamine or amine. Acyl sulfonamide 111 can besynthesized by preparing an activated ester of carboxylic acid 109followed by treatment with the appropriate sulfonamide and tertiaryamine base in a suitable organic solvent such as THF. In one embodiment,acyl sulfonamide 111 is prepared by treatment of carboxylic acid 109with CDI at elevated temperature (50° C.) in THF followed by treatmentwith the appropriate sulfonamide and DBU.

Scheme 2 illustrates an alternative method for synthesizing compounds ofthe Formula I. Nitrile 113 can be prepared by palladium mediatedcoupling of bromide 104 with zinc cyanide in suitable organic solventsuch as DMA, NMP or DMF at elevated temperatures ranging from 50 to 120°C. Several palladium catalysts may be employed including but not limitedto Pd(PPh₃)₄, PdCl₂(dppf), or Pd₂ dba₃ with ligands such as dppe, dppp,dppf or BINAP. In one embodiment, nitrile 113 is prepared from bromide104 by treatment with zinc cyanide, Pd₂ dba₃, and dppf in NMP at 120° C.Amino benzisoxazole 114 can be prepared in a two step procedure fromnitrile 113 by the addition of the potassium salt of acetone oxime insuitable organic solvent such as THF or Et₂O at temperatures rangingfrom −78 to 5° C. followed by acid catalyzed cyclization. In oneembodiment the acetone oxime addition can be performed by addition of aTHF solution of nitrile 113 to the salt at 0° C. in THF followed bywarming to room temperature. The cyclization can be accomplished under avariety of acidic conditions at a range of temperatures. In oneembodiment, the cyclization method comprises treatment of the oximeaddition product in MeOH with 2M HCl in Et₂O. Halogenation to formbenzisoxazole 115 is accomplished using standard procedures such as NCSor NBS in DMF. Compound 116 is prepared in a two step procedurecomprising hydrolysis of ester 115 under standard conditions to form thecorresponding carboxylic acid, followed by conversion of the carboxylicacid to hydroxamate 116 by standard coupling procedures, including butnot limited, to EDCI/HOBt, PyBOP, or DIC and the appropriatehydroxylamine.

Scheme 3 illustrates a method of synthesizing compounds of the FormulaII. 4,6-Dichloronicotinic acid 118 can be prepared from4,6-dihydroxynicotinic acid ethyl ester 117 in two steps. In the firststep, 4,6-dihydroxynicotinic acid ethyl ester 117 is chlorinated usingan appropriate reagent such as POCl₃, oxalyl chloride or thionylchloride. In one embodiment, chlorination is accomplished with POCl₃ andEt₃N at elevated temperatures. Hydrolysis of the resulting dichloroethylester to provide compound 118 can be performed under standardconditions. Aniline 119 can be prepared by deprotonation of the properlysubstituted aniline with KHMDS, LiHMDS, NaHMDS or LDA at low temperature(−100 to −60° C.) in appropriate ethereal solvent such as THF or diethylether followed by addition of carboxylic acid 118 and warming to roomtemperature. In one embodiment, deprotonation is accomplished withLiHMDS at −78° C. in THF, followed by addition of carboxylic acid 118and warming to room temperature. Aminopyridine 120 is prepared in threesteps from aniline 119. In the first step, the tert-butyl ester isprepared by treating the acid 119 with2-tert-butyl-1,3-diisopropylisourea in THF at temperatures ranging from25 to 75° C. In the second step, sodium azide is added to the tert-butylester in DMF at 80° C. The aminopyridine 120 is prepared by reduction ofthe azide under standard conditions including but not limited to Zndust/AcOH, Pt/C or PtO₂ in the presence of H₂ gas, Ph₃P or SnCl₂/MeOH.In one embodiment, the azide reduction is accomplished by treatment withZn dust in a mixture of methylene chloride and acetic acid.Imidazopyridine 121 wherein Z=F is prepared in two steps fromaminopyridine 120. In the first step, fluorination is accomplished bytreatment of the aminopyridine 120 with SELECTFLUOR® in a mixture ofMeOH and water or pH 7 phosphate buffer. Cyclization to formimidazopyridine 121 (Z=H or F) can be accomplished by treatment withchloroacetaldehyde or bromoacetaldehyde in suitable organic solvent suchas DMF or EtOH at elevated temperatures (50 to 120° C.). In oneembodiment, cyclization is realized by treatment with chloroacetaldehydein EtOH at 70° C. Alternatively, aniline 119 can be converted todichloroester 122 in two steps. In the first step, chlorination isperformed under standard conditions such as NCS in DMF. In the secondstep, esterification can be achieved by standard methods including butnot limited to Fisher esterification (MeOH, H₂SO₄), reaction withTMSCHN₂ or TMSCI in MeOH. Aminopyridine 123 can be prepared as describedabove for aminopyridine 120 with the exception that the sodium azideaddition can be accomplished at room temperature. Cyclization (achievedas described above for imidazopyridine 121) followed by standard basicsaponification gives imidazopyridine 124. Hydroxamate 125 can beprepared from either imidazopyridine 121 or 124 using standard couplingprocedures including but not limited to EDCI/HOBt, PyBOP, or DIC and theappropriate hydroxylamine. Alternatively, hydroxamate 125 can beprepared in two steps by initial conversion to the acid chloride bystandard methods followed by addition of the hydroxylamine.

Scheme 4 illustrates an alternative method of preparing compounds ofFormula II. An appropriately functionalized 2-aminopyridine 126 in asuitable organic solvent such as dichloromethane or dichloroethane isreacted with a Lewis acid such as zinc bromide and condensation product127 as disclosed by Katritzky et al. (J. Org. Chem., 2003, 68,4935-4937: J. Org. Chem., 1990, 55, 3209-3213) to provide the3-dialkyamino-imidazo[1,2-a]pyridine ring system 128. Condensationproducts 127 (i.e., condensation of a glyoxal, benzotriazole and asecondary amine) can be generated using benzotriazole, glyoxal and anyappropriate secondary amine including, but not limited to dimethylamine,diethylamine, pyrrolidine, piperidine, morpholine, 1-methylpiperazine,N-methylallylamine, diallyamine, and N-methylbenzylamine. The ester 128is hydrolyzed by standard saponification methods, and the resulting acidcan be converted to hydroxamate 129 by standard coupling proceduresincluding but not limited to EDCI/HOBt, PyBOP, or DIC and theappropriate hydroxylamine. Alternatively, hydroxamate 129 can beprepared in two steps by initial conversion of the carboxylic acid tothe acid chloride or activated ester by standard methods followed byaddition of the hydroxylamine.

Scheme 5 illustrates an alternative method of preparing compounds of theFormula II. The preparation of 3-aminomethylimidazo[1,2-a]pyridines 131using the modified Mannich reaction procedure developed by Kercher etal. (manuscript in preparation) is illustrated. The reaction isgenerally carried out by combining 37% aqueous formaldehyde and asuitable amine in 6:1 acetonitrile/water. Several secondary amines canbe employed including but not limited to pyrrolidine, piperidine,morpholine, dimethylamine, N-BOC-piperazine and 1-methylpiperazine. Thesolution of amine and formaldehyde is stirred for approximately half anhour after which time scandium triflate and the appropriateimidazo[1,2-a]pyridine 130 are sequentially added. In one embodiment,the Mannich reaction is catalyzed by a Group IIIA lanthanide triflate,such as scandium triflate, though alternatively it may be performedusing an excess of protic acid (AcOH or HCl) or elevated temperatures.

Scheme 6 illustrates an alternative method of preparing compounds ofFormula II. In Scheme 6, the preparation of3-aminomethylimidazo[1,2-a]pyridines 134 via reductive alkylation isillustrated. The 3-aminomethylimidazo[1,2-a]pyridine 133 is preparedfrom the appropriate 3-formylimidazo[1,2-a]pyridine 132 and a suitableamine using standard reduction methods such as Na(CN)BH₃, Na(OAc)₃BH,NMe₄BH(OAc)₃ with or without the addition of acetic acid in a suitablenonreactive organic solvent such as methylene chloride, acetonitrile ortetrahydrofuran. The reductive amination is generally accomplished bytreatment of the aldehyde derivative 132 with the amine and acetic acidin tetrahydrofuran at room temperature followed by the addition ofNa(OAc)₃BH. In cases wherein R″=H, the corresponding secondary amine 133can optionally be protected, for example with an acid labile protectinggroup such as tert-butyl carbamate (BOC) to facilitate handling insubsequent steps. The ester is hydrolyzed by standard saponificationmethods, and the resulting acid can be converted to hydroxamate 134 bystandard coupling procedures including but not limited to EDCI/HOBt,PyBOP, or DIC and the appropriate hydroxylamine. Alternatively,hydroxamate 134 can be prepared in two steps by initial conversion ofthe carboxylic acid to the acid chloride or activated ester by standardmethods followed by addition of the hydroxylamine. Protecting groups, ifpresent, are removed after coupling.

Scheme 7 illustrates a method of preparing compounds of Formula III. InScheme 7 the preparation of 3-alkyl-[1,2,4]triazolo[4,3-a]pyridinederivatives is illustrated. Compound 136 is prepared from compound 135in a two-step process. A suitably functionalized 2-chloropyridinederivative 135 is converted to the 2-hydrazinopyridine by reaction withhydrazine. The reaction is generally accomplished by reaction ofhydrazine with the 2-chloropyridine derivative 135 in an unreactiveorganic solvent such as DMF or DMA at elevated temperature (50 to 100°C.). The 2-hydrazinopyridine is then acylated with the appropriatecarboxylic acid halide such as fluoride, chloride or bromide, or theappropriate carboxylic acid anhydride or mixed anhydride in a suitableunreactive organic solvent such as dichloromethane, and in the presenceof a suitable base such as triethylamine, diisopropylethylamine orpyridine, to provide intermediate 136. Acylation of the2-hydrazinopyridine can alternatively be accomplished by standardpeptide coupling procedures with the appropriate carboxylic acid andappropriate coupling reagent, including but not limited to EDCI/HOBt,PyBOP, or DIC. The intermediate 136 is converted to3-alkyl-[1,2,4]triazolo[4,3-a]pyridine 137 by treatment with an excessof phosphorus oxychloride in refluxing dichloromethane. The ester 137 ishydrolyzed by standard saponification methods, and the resulting acid138 can be converted to hydroxamate 139 by standard peptide couplingprocedures including but not limited to EDCI/HOBt, PyBOP, or DIC and theappropriate hydroxylamine. Alternatively, hydroxamate 139 can beprepared in two steps by initial conversion of the carboxylic acid tothe acid chloride or activated ester by standard methods followed byaddition of the hydroxylamine.

Scheme 8 illustrates a method of preparing compounds of the Formula IV.In Scheme 8, the synthesis of 3-methyl-benzo[c]isoxazole derivatives isillustrated. Compound 141 is prepared from compound 140 in a two-stepprocess. Methyl ester 140 is treated with sodium azide in 3:1acetone/water at elevated temperature (reflux) to effect nucleophilicsubstitution. The 4-azido derivative is then isolated and heated inwater at reflux to effect cyclization to the benzo[c]isoxazole ringsystem 141. The ester 141 is hydrolyzed by standard saponificationmethods, and the resulting carboxylic acid can be converted tohydroxamate 142 by standard peptide coupling procedures including butnot limited to EDCI/HOBt, PyBOP, or DIC and the appropriatehydroxylamine. Alternatively, hydroxamate 142 can be prepared in twosteps by initial conversion of the carboxylic acid to the acid chlorideor activated ester by standard methods followed by addition of thehydroxylamine.

Scheme 9 illustrates a method of preparing compounds of Formula V.2-Chloro-4-methyl-5-nitropyridine 143 can be converted to amino pyridine144 in a three-step sequence. In the first step, Sonagashria couplingusing TMS-acetylene, CuI, amine base, palladium catalyst and organicsolvent such as DME, THF, or DMF at temperatures from 25 to 100° C.gives the nitroacetylenic pyridine. Suitable palladium catalystsinclude, but are not limited to, PdCl₂(dppf), Pd(Ph₃P)₄, Pd(PPh₃)₂Cl₂and Pd₂ dba₃/dppf. Suitable amine bases include, but are not limited to,Et₃N, Hunig's base, and diisopropylamine. The amino pyridine 144 is thenprepared by removal of the TMS group under standard conditions such asK₂CO₃ in MeOH, followed by reduction of the nitro group using either Zndust/AcOH, Fe or SnCl₂/MeOH. For Z=H, aminopyridine 144 is used directlyin the cyclization reaction. When Z=Cl, aminopyridine 144 is halogenatedunder standard conditions with NCS in DMF and then carried forward tothe cyclization. When Z=F, the 2-chloro-3-aminopyridine intermediate istreated with KF, Kryptofix in DMSO to prepare amino pyridine 145.Cyclization to give pyrazolo[1,5-a]pyridine 146 is accomplished bytreating aminopyridine 145 with O-(4-nitrophenyl)-hydroxylamine in asuitable organic solvent such as DMF at room temperature in presence ofa base such as K₂CO₃. Carboxylic acid 149 can be prepared, for example,using one the following routes. One route involves palladium mediatedcross-coupling with appropriately substituted bromobenzene andaminopyrazolo[1,5-a]pyridine 146. In this case, the cross-coupling canbe accomplished with palladium catalyst and organic solvent such as DME,THF, dioxane, and toluene at temperatures from 60 to 120° C. Suitablepalladium catalysts include, but are not limited to, Pd(OAc)₂,PdCl₂(dppf), Pd₂(bda)₃, and Pd(dba)₂. Suitable ligands include, but arenot limited to, BINAP, DPPF, and (o-tol)₃P. Suitable amine basesinclude, but are not limited to, NaOt-Bu, KOt-Bu, and Cs₂CO₃. The secondroute involves S_(N)Ar reaction with aminopyrazolo[1,5-a]pyridine 146and the appropriately substituted 2-fluoronitrobenzene. In this case,the coupling can be accomplished by mixing the two components in asuitable organic solvent such as xylenes, toluene, DMSO or DMF atelevated temperatures (80 to 150° C.). Optionally, a base can beemployed in the S_(N)Ar coupling such as K₂CO₃ or Cs₂CO₃. The carboxylicacid 149 is then prepared by functionalization of the aromatic ringfollowed by oxidation. In the first case, functionalization involveshalogenation under standard conditions with either NCS or NBS in DMF. Inthe second case, functionalization involves Sandmeyer chemistry toconvert the nitroarene into the desired arene or arylhalide (nitro groupreduction; diazonation; halogentation or protonation). In both routes,the last step to prepare carboxylic acid 149 is oxidation of the toluylmoiety. This can be achieved using standard methods including but notlimited to KMnO₄, NaOCl/RuCl₃ or Na₂Cr₂O₇/HCl. The resulting carboxylicacid 149 can be converted to hydroxamate 150 by standard peptidecoupling procedures including but not limited to EDCIIHOBt, PyBOP, orDIC and the appropriate hydroxylamine. Alternatively, hydroxamate 150can be prepared in two steps by initial conversion of the carboxylicacid to the acid chloride or activated ester by standard methodsfollowed by addition of the hydroxylamine.

The present invention provides methods for treating chronic inflammatorydiseases. The method comprises administering an effective amount of acompound of Formula I-V, or a pharmaceutically acceptable salt, prodrug,solvate or pharmaceutical composition thereof, to a mammal, such as ahuman, in need of such treatment.

As used herein, the term “inflammatory disease” and “inflammatorydisorder” includes a disease or disorder characterized by, caused by,resulting from, or becoming affected by inflammation. Examples ofinflammatory diseases or disorders include, but not limited to, acuteand chronic inflammation disorders such as rheumatoid arthritis,osteoarthritis, inflammatory bowel diseases (including, but not limitedto, Crohn's disease and ulcerative colitis), chronic obstructivepulmonary disorder (COPD), psoriasis, multiple sclerosis, asthma,diseases and disorders related to diabetic complications, fibrotic organfailure in organs such as lung, liver, kidney, and inflammatorycomplications of the cardiovascular system such as acute coronarysyndrome.

The compounds of Formulas I-V may also be used in the treatment ofinflammatory diseases in combination with one or more additional agentssuch as non-steroidal anti-inflammatory agents (NSAIDs), such asibuprofen or aspirin (which reduce swelling and alleviate pain);disease-modifying anti-rheumatic drugs (DMARDs) such as methotrexate;5-aminosalicylates (sulfasalazine and the sulfa-free agents);corticosteroids; immunomodulators such as 6-mercaptoputine (“6-MP”),azathioprine (“AZA”), cyclosporines, and biological response modifierssuch as REMICADE (infliximab) and ENBREL (etanercept); fibroblast growthfactors; platelet derived growth factors; enzyme blockers such as ARAVA(leflunomide); and/or a cartilage protecting agent such as hyaluronicacid, glucosamine, chondroitin, etc.

The term “treating,” as used herein, unless otherwise indicated, meansreversing, alleviating, inhibiting the progress of, or preventing thedisorder or condition to which such term applies, or one or moresymptoms of such disorder or condition, and includes, but is not limitedto, preventing the disease condition from occurring in a mammal,particularly when the mammal is found to be predisposed to having thedisease condition but has not yet been diagnosed as having it;modulating and/or inhibiting the disease condition; and/or alleviatingthe disease condition. The term “treatment,” as used herein, unlessotherwise indicated, refers to the act of treating as “treating” isdefined immediately above.

The amount of a given agent that will correspond to such an amount willvary depending upon factors such as the particular compound, diseasecondition and its severity, the identity (e.g., weight) of the mammal inneed of treatment, but can nevertheless be routinely determined by oneskilled in the art.

In order to use a compound of the Formula I-V or a solvate, metabolite,pharmaceutically acceptable salt or prodrug thereof, for the therapeutictreatment (including prophylactic treatment) of mammals includinghumans, it is normally formulated in accordance with standardpharmaceutical practice as a pharmaceutical composition. According tothis aspect of the invention there is provided a pharmaceuticalcomposition that comprises a compound of the Formula I-V, or apharmaceutically acceptable salt or prodrug thereof, in association witha pharmaceutically acceptable diluent or carrier.

Accordingly, the invention also includes a method of treating ahyperproliferative disorder in a mammal, wherein the method comprisesadministering to said mammal a composition comprising an effectiveamount of one or more compounds of Formula I-V, or a solvate,metabolite, pharmaceutically acceptable salt or prodrug thereof, and apharmaceutically acceptable carrier.

The compositions of the invention may be in a form suitable for oral use(for example as tablets, lozenges, hard or soft capsules, aqueous oroily suspensions, emulsions, dispersible powders or granules, syrups orelixirs), for topical use (for example as creams, ointments, gels, oraqueous or oily solutions or suspensions), for administration byinhalation (for example as a finely divided powder or a liquid aerosol),for administration by insufflation (for example as a finely dividedpowder) or for parenteral administration (for example as a sterileaqueous or oily solution for intravenous, subcutaneous, or intramusculardosing or as a suppository for rectal dosing). For example, compositionsintended for oral use may contain, for example, one or more coloring,sweetening, flavoring and/or preservative agents.

Suitable pharmaceutically-acceptable excipients for a tablet formulationinclude, for example, inert diluents such as lactose, sodium carbonate,calcium phosphate or calcium carbonate, granulating and disintegratingagents such as corn starch or algenic acid; binding agents such asstarch; lubricating agents such as magnesium stearate, stearic acid ortalc; preservative agents such as ethyl or propyl p-hydroxybenzoate, andanti-oxidants, such as ascorbic acid. Tablet formulations may beuncoated or coated either to modify their disintegration and thesubsequent absorption of the active ingredient within thegastrointestinal tract, or to improve their stability and/or appearance,in either case, using conventional coating agents and procedures wellknown in the art.

Compositions for oral use may be in the form of hard gelatin capsules inwhich the active ingredient is mixed with an inert solid diluent, forexample, calcium carbonate, calcium phosphate or kaolin, or as softgelatin capsules in which the active ingredient is mixed with water oran oil such as peanut oil, liquid paraffin, or olive oil.

Aqueous suspensions generally contain the active ingredient in finelypowdered form together with one or more suspending agents such as sodiumcarboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose,sodium alginate, polyvinyl-pyrrolidone, gum tragacanth and gum acacia;dispersing or wetting agents such as lecithin or condensation productsof an alkylene oxide with fatty acids (for example polyoxethylenestearate), or condensation products of ethylene oxide with long chainaliphatic alcohols, for example heptadecaethyleneoxycetanol, orcondensation products of ethylene oxide with partial esters derived fromfatty acids and a hexitol such as polyoxyethylene sorbitol monooleate,or condensation products of ethylene oxide with partial esters derivedfrom fatty acids and hexitol anhydrides, for example polyethylenesorbitan monooleate. The aqueous suspensions may also contain one ormore preservatives (such as ethyl or propyl p-hydroxybenzoate,anti-oxidants (such as ascorbic acid), coloring agents, flavoringagents, and/or sweetening agents (such as sucrose, saccharine oraspartame).

Oily suspensions may be formulated by suspending the active ingredientin a vegetable oil (such as arachis oil, olive oil, sesame oil orcoconut oil) or in a mineral oil (such as liquid paraffin). The oilysuspensions may also contain a thickening agent such as beeswax, hardparaffin or cetyl alcohol. Sweetening agents such as those set outabove, and flavoring agents may be added to provide a palatable oralpreparation. These compositions may be preserved by the addition of ananti-oxidant such as ascorbic acid.

Dispersible powders and granules suitable for preparation of an aqueoussuspension by the addition of water generally contain the activeingredient together with a dispersing or wetting agent, suspending agentand one or more preservatives. Suitable dispersing or wetting agents andsuspending agents are exemplified by those already mentioned above.Additional excipients such as sweetening, flavoring and coloring agents,may also be present.

The pharmaceutical compositions of the invention may also be in the formof oil-in-water emulsions. The oily phase may be a vegetable oil, suchas olive oil or arachis oil, or a mineral oil, such as for exampleliquid paraffin or a mixture of any of these. Suitable emulsifyingagents may be, for example, naturally-occurring gums such as gum acaciaor gum tragacanth, naturally-occurring phosphatides such as soya bean,lecithin, esters or partial esters derived from fatty acids and hexitolanhydrides (for example sorbitan monooleate) and condensation productsof the said partial esters with ethylene oxide such as polyoxyethylenesorbitan monooleate. The emulsions may also contain sweetening,flavoring and preservative agents.

Syrups and elixirs may be formulated with sweetening agents such asglycerol, propylene glycol, sorbitol, aspartame or sucrose, and may alsocontain a demulcent, preservative, flavoring and/or coloring agent.

The pharmaceutical compositions may also be in the form of a sterileinjectable aqueous or oily suspension, which may be formulated accordingto known procedures using one or more of the appropriate dispersing orwetting agents and suspending agents, which have been mentioned above. Asterile injectable preparation may also be a sterile injectable solutionor suspension in a non-toxic parenterally acceptable diluent or solvent,for example a solution in 1,3-butanediol.

Suppository formulations may be prepared by mixing the active ingredientwith a suitable non-irritating excipient which is solid at ordinarytemperatures but liquid at the rectal temperature and will thereforemelt in the rectum to release the drug. Suitable excipients include, forexample, cocoa butter and polyethylene glycols.

Topical formulations, such as creams, ointments, gels and aqueous oroily solutions or suspensions, may generally be obtained by formulatingan active ingredient with a conventional, topically acceptable, vehicleor diluent using conventional procedures well known in the art.

Compositions for administration by insufflation may be in the form of afinely divided powder containing particles of average diameter of, forexample, 30 μm or much less, the powder itself comprising either activeingredient alone or diluted with one or more physiologically acceptablecarriers such as lactose. The powder for insufflation is thenconveniently retained in a capsule containing, for example, 1 to 50 mgof active ingredient for use with a turbo-inhaler device, such as isused for insufflation of the known agent sodium cromoglycate.

Compositions for administration by inhalation may be in the form of aconventional pressurized aerosol arranged to dispense the activeingredient either as an aerosol containing finely divided solid orliquid droplets. Conventional aerosol propellants such as volatilefluorinated hydrocarbons or hydrocarbons may be used and the aerosoldevice is conveniently arranged to dispense a metered quantity of activeingredient.

For further information on formulations, see Chapter 25.2 in Volume 5 ofComprehensive Medicinal Chemistry (Corwin Hansch; Chairman of EditorialBoard), Pergamon Press 1990, which is specifically incorporated hereinby reference.

The amount of a compound of this invention that is combined with one ormore excipients to produce a single dosage form will necessarily varydepending upon the subject treated, the severity of the disorder orcondition, the rate of administration, the disposition of the compoundand the discretion of the prescribing physician. For example, aneffective dosage may be in the range of about 1 to about 60 mg per kgbody weight per day, in single or divided doses. In some instances,dosage levels below the lower limit of the aforesaid range may be morethan adequate, while in other cases still larger doses may be employedwithout causing any harmful side effect, provided that such larger dosesare first divided into several small doses for administration throughoutthe day. For further information on routes of administration and dosageregimes, see Chapter 25.3 in Volume 5 of Comprehensive MedicinalChemistry (Corwin Hansch; Chairman of Editorial Board), Pergamon Press1990, which is specifically incorporated herein by reference.

The size of the dose for therapeutic or prophylactic purposes of acompound of Formula I-V will naturally vary according to the nature andseverity of the conditions, the age and sex of the animal or patient andthe route of administration, according to well known principles ofmedicine.

The activity of the compounds of the present invention may be determinedby the following procedure. N-terminal 6 His-tagged, constitutivelyactive MEK-1 (2-393) is expressed in E. coli and protein is purified byconventional methods (Ahn, et al., Science 1994, 265, 966-970). Theactivity of MEK1 is assessed by measuring the incorporation ofγ-³³P-phosphate from γ-³³P-ATP onto N-terminal His tagged ERK2, which isexpressed in E. coli and is purified by conventional methods, in thepresence of MEK-1. The assay is carried out in 96-well polypropyleneplate. The incubation mixture (100 μL) comprises of 25 mM Hepes, pH 7.4,10 mM MgCl₂, 5 mM O-glycerolphosphate, 100 μM Na-orthovanadate, 5 mMDTT, 5 nM MEK1, and 1 μM ERK2. Inhibitors are suspended in DMSO, and allreactions, including controls are performed at a final concentration of1% DMSO. Reactions are initiated by the addition of 10 VtM ATP (with 0.5μCi γ-³³P-ATP/well) and incubated at ambient temperature for 45 minutes.Equal volume of 25% TCA is added to stop the reaction and precipitatethe proteins. Precipitated proteins are trapped onto glass fiber Bfilterplates, and excess labeled ATP washed off using a Tomtec MACH IIIharvester. Plates are allowed to air-dry prior to adding 30 μL/well ofPackard Microscint 20, and plates are counted using a Packard TopCount.In this assay, compounds of the invention exhibited an IC₅₀ of less than50 micromolar.

The examples presented below are intended to illustrate particularembodiments of the invention, and are not intended to limit the scope ofthe specification or the claims in any way.

CHEMICAL EXAMPLES

In order to illustrate the invention, the following examples areincluded. However, it is to be understood that these examples do notlimit the invention and are only meant to suggest a method of practicingthe invention. Persons skilled in the art will recognize that thechemical reactions described may be readily adapted to prepare a numberof other MEK inhibitors of the invention, and alternative methods forpreparing the compounds of this invention are deemed to be within thescope of this invention. For example, the synthesis of non-exemplifiedcompounds according to the invention may be successfully performed bymodifications apparent to those skilled in the art, e.g., byappropriately protecting interfering groups, by utilizing other suitablereagents known in the art other than those described, and/or by makingroutine modifications of reaction conditions. Alternatively, otherreactions disclosed herein or known in the art will be recognized ashaving applicability for preparing other compounds of the invention.

In the examples described below, unless otherwise indicated alltemperatures are set forth in degrees Celsius. Reagents were purchasedfrom commercial suppliers such as Aldrich Chemical Company, Lancaster,TCI or Maybridge, and were used without further purification unlessotherwise indicated. Tetrahydrofuran (THF), N,N-dimethylformamide (DMF),dichloromethane, toluene, and dioxane were purchased from Aldrich inSure seal bottles and used as received.

The reactions set forth below were done generally under a positivepressure of nitrogen or argon or with a drying tube (unless otherwisestated) in anhydrous solvents, and the reaction flasks were typicallyfitted with rubber septa for the introduction of substrates and reagentsvia syringe. Glassware was oven dried and/or heat dried.

Column chromatography was done on a Biotage system (Dyax Corporation)having a silica gel column or on a silica SepPak cartridge (Waters).

¹H-NMR spectra were recorded on a Varian instrument operating at 400MHz. ¹H-NMR spectra were obtained as CDCl₃, CD₃OD or DMSO-d₆ solutions(reported in ppm), using chloroform as the reference standard (7.25ppm). Other NMR solvents were used as needed. When peak multiplicitiesare reported, the following abbreviations are used: s (singlet), d(doublet), t (triplet), m (multiplet), br (broadened), dd (doublet ofdoublets), dt (doublet of triplets). Coupling constants, when given, arereported in Hertz (Hz).

Example 1

Synthesis of6-(4-bromo-2-chlorophenylamino)-7-fluoro-3-methylbenzo[d]isoxazole-5-carboxylicacid (9a)

Step A: Preparation of 5-bromo-2,3,4-trifluorobenzoic acid (2): To asolution of 1-bromo-2,3,4-trifluorobenzene (1) (5.0 mL, 41.7 mmol) inTHF (120 mL) was added LiHMDS (2.0 M solution, 21 mL, 42 mmol) at −78°C. After stirring for 1 hour at −78° C., the mixture was added to asolution of CO₂ in THF (1 L). The dry-ice bath was removed and thereaction mixture stirred overnight at room temperature. The reactionmixture was quenched with 10% aqueous HCl (835 mL), concentrated, andwashed with ether (250 mL). The combined organics were washed with 5%aqueous NaOH (300 mL) and water (100 mL). The aqueous layer wasacidified (pH 0) with concentrated HCl. The resulting suspension wasextracted with ether (2×300 mL), dried over MgSO₄, filtered,concentrated under reduced pressure to afford 7.70 g (72% yield) of thedesired product (2).

Step B: Preparation of5-bromo-2-(2-chlorophenylamino)-3,4-difluorobenzoic acid (3): To asolution of LiHMDS (49.0 mL, 2 M in THF/heptane) in THF (40 mL) wasadded 2-chlorophenylamine (6.50 mL, 60.6 mmol) at −78° C. After vigorousstirring for 10 minutes, a solution of 5-bromo-2,3,4-trifluoro-benzoicacid (2) (7.70 g, 30.2 mmol) in THF (60 mL) was added. The dry-ice bathwas removed and the reaction mixture stirred for 4 hours at roomtemperature. The mixture was concentrated, treated with 10% aqueous HCl(75 mL), and extracted with EtOAc. The combined organic extracts weredried over MgSO₄, filtered, and concentrated. Purification bytrituration with boiling CH₂Cl₂ gave 7.24 g (66%) of the desired acid(3) as a yellow solid

Step C: Preparation of5-bromo-2-(2-chlorophenylamino)-3,4-difluorobenzoic acid methyl ester(4): To a solution of5-bromo-2-(2-chlorophenylamino)-3,4-difluorobenzoic acid (3) (4.50 g,12.4 mmol) in a 3:1 mixture of THF:MeOH (32 mL) was addedtrimethylsilyldiazomethane (8.10 ml of a 2 M solution in hexanes) atroom temperature. After stirring for 2 hours, the reaction mixture wasquenched with acetic acid, diluted with EtOAc, and washed with water.The organic layer was dried (MgSO₄) and concentrated under reducedpressure to give 4.35 g (93%) of the desired methyl ester (4).

Step D: Preparation of2-(2-chlorophenylamino)-3,4-difluoro-5-trimethylsilanylethynylbenzoicacid methyl ester (5): A mixture of5-bromo-2-(2-chlorophenylamino)-3,4-difluorobenzoic acid methyl ester(4) (101 mg, 0.268 mmol), TMS-acetylene (0.045 mL, 0.31 mmol),Pd(PPh₃)₂Cl₂ (18.7 mg, 0.0261 mmol), CuI (5.1 mg, 0.027 mmol), andi-Pr₂NH (0.075 mL, 0.53 mmol) in THF (1.5 mL) was stirred for 16 hoursat room temperature. The reaction mixture was concentrated under reducedpressure, and diluted with EtOAc. The organic layer was washed withsaturated aqueous NH₄Cl and brine, dried over MgSO₄, and concentrated.Purification by flash column chromatography using the Biotage system(100% hexane to 1% EtOAc in hexane) gave 81.3 mg (77% yield) of thedesired product (5).

Step E: Preparation of5-acetyl-2-(2-chlorophenylamino)-3,4-difluorobenzoic acid methyl ester(6): A mixture of2-(2-chlorophenylamino)-3,4-difluoro-5-trimethylsilanylethynylbenzoicacid methyl ester (5) (79.4 mg, 0.20 mmol), HgSO₄ (59.8 mg, 2.0 mmol),and conc. H₂SO₄ (0.02 mL, 0.40 mmol) in 80% aqueous acetone (2.5 mL),were refluxed for 48 hours. The reaction was concentrated under reducedpressure, and diluted with EtOAc. The organic layer was washed withwater and brine, dried over MgSO₄ and concentrated to give 50.1 mg (73%)of the desired product (6).

Step F: Preparation of6-(2-chlorophenylamino)-7-fluoromethylbenzo[d]isoxazole-5-carboxylicacid methyl ester (7): t-BuOK (0.47 mL, 1.0 M in THF) was added topropan-2-one oxime (35 mg, 0.47 mmol). After stirring for 30 minutes,THF (0.5 mL) was added, and the reaction mixture was cooled to −78° C. Asolution of 5-acetyl-2-(2-chlorophenylamino)-3,4-difluorobenzoic acidmethyl ester (6) (50.0 mg, 0.147 mmol) in THF (1 mL) was added. Thereaction mixture was slowly warmed to 0° C. and stirred for 2 hours. Thereaction mixture was quenched with saturated aqueous NH₄Cl, diluted withEtOAc and water. The aqueous layer was separated and extracted withEtOAc. The combined organic extracts were dried over MgSO₄, filtered,and concentrated in vacuo to give5-acetyl-2-(2-chlorophenylamino)-3-fluoro-4-isopropylideneaminooxybenzoicacid methyl ester. The recovered oxime was suspended in a 1:1 mixture of5% aqueous HCl and MeOH (30 mL) and heated to reflux. After 1 hour, thereaction mixture was cooled to room temperature and diluted with EtOAc.The organic layer was washed with water, dried (MgSO₄) and concentrated.Purification by flash column chromatography using the Biotage system(40% methylene chloride in hexanes) provided 17 mg (35% for two steps)of the desired product (7).

Step G: Preparation of6-(4-bromo-2-chlorophenylamino)-7-fluoro-3-methylbenzo[d]isoxazole-5-carboxylicacid methyl ester (8a):6-(2-Chlorophenylamino)-7-fluoro-3-methylbenzo[d]isoxazole-5-carboxylicacid methyl ester (7) (18.6 mg, 0.0556 mmol) and N-bromosuccinimide(12.0 mg, 0.0667 mmol) were stirred in DMF (1 mL) for 16 hours. Thereaction mixture was diluted with EtOAc, and washed with water (2×). Theorganic layer was dried over MgSO₄, filtered, and concentrated.Purification by flash column chromatography using the Biotage system(10% EtOAc in hexanes) provided 12.6 mg (55%) of the desired product(8a).

Step H: Preparation of6-(4-bromo-2-chlorophenylamino)-7-fluoro-3-methylbenzo[d]isoxazole-5-carboxylicacid (9a): To a solution of6-(4-bromo-2chlorophenylamino)-7-fluoro-3-methylbenzo[d]isoxazole-5-carboxylicacid methyl ester (8a) (200 mg, 0.48 mmol) in THF-water (3 mL/1.5 mL)was added aqueous LiOH (1 M, 1.00 mL) at room temperature. After 15hours, the reaction mixture was acidified to pH 1 with aqueous HCl (1M), diluted with water, and extracted with EtOAc/THF. The organic layerwas washed with water, dried over MgSO₄, filtered, and concentrated invacuo to give 191.5 mg (99%) of the crude acid (9a) which was usedwithout further purification. MS APCI (−) m/z 397, 399 (M+, Br, Clpattern) detected. ¹H NMR (400 MHz, DMSO-d₆) δ 9.55 (s, 1H), 8.37 (s,1H), 7.75 (s, 1H), 7.42 (d, 1H), 6.97 (t, 1H), 2.60 (s, 3H): ¹⁹F NMR(376 MHz, DMSO-d₆) −140.15 (s).

Example 2

Synthesis of6-(4-bromo-2-chlorophenylamino)-7-fluoro-3-methylbenzo[d]isoxazole-5-carboxylicacid cyclopropylmethoxyamide (10a)

To a solution of6-(4-bromo-2-chlorophenylamino)-7-fluoro-3-methylbenzo[d]isoxazole-5-carboxylicacid (9a) (50.0 mg, 0.125 mmol) in DMF (1 mL) was added HOBt (24.6 mg,0.161 mmol), Et₃N (0.060 mL, 0.43 mmol),O-cyclopropylmethyl-hydroxylamine (15.5 mg, 0.178 mmol), and1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDCI) (32.2mg, 0.168 mmol) at room temperature. After 6 days, the reaction mixturewas diluted with EtOAc, washed with saturated aqueous NH₄Cl, brine,saturated aqueous NaHCO₃, and brine. The organic layer was dried overMgSO₄, filtered, concentrated in vacuo, and purified by flash columnchromatography using the Biotage system (0.5% MeOH in CH₂Cl₂) to give27.6 mg (47% yield) of the desired product (10a). MS APCI (−) m/z 466,468 (M+, Br, Cl pattern) detected. ¹H NMR (400 MHz, CD₃OD) δ 7.82 (s,1H), 7.57 (d, 1H), 7.31 (dd, 1H), 6.74 (dd, 1H), 3.73 (d, 2H), 2.60 (s,3H), 1.16 (m, 1H), 0.55 (m, 2H), 0.28 (m, 2H). ¹⁹F NMR (376 MHz,CD₃OD)−140.96 (s, 1F).

Example 3

Synthesis of6-(4-bromo-2-chlorophenylamino)-7-fluoro-3-methylbenzo[d]isoxazole-5-carboxylicacid (2-hydroxyethoxy)-amide (12a)

Step A: Preparation of6-(4-bromo-2-chlorophenylamino)-7-fluoro-3-methyl-benzo[d]isoxazole-5-carboxylicacid (2-vinyloxyethoxy)-amide (11a): To a solution of6-(4-bromo-2-chlorophenylamino)-7-fluoro-3-methylbenzo[d]isoxazole-5-carboxylicacid (10a) (75.2 mg, 0.188 mmol) in DMF (1.5 mL) was added HOBt (38.2mg, 0.249 mmol), Et₃N (0.080 mL, 0.571 mmol),O-(2-vinyloxyethyl)hydroxylamine (28.5 mg, 0.276 mmol), and EDCI (47.2mg, 0.246 mmol) at room temperature. After 6 days, the reaction mixturewas diluted with EtOAc, washed with saturated aqueous NH₄Cl, brine,saturated aqueous NaHCO₃, and brine. The organic layer was dried overMgSO₄, filtered, concentrated in vacuo, and purified by flash columnchromatography using the Biotage system (3% MeOH in CH₂Cl₂) to give 57.8mg (63%) of the desired product (11a).

Step B: Preparation of6-(4-bromo-2-chlorophenylamino)-7-fluoro-3-methylbenzo[d]isoxazole-5-carboxylicacid (2-hydroxyethoxy)-amide (12a): A solution of6-(4-bromo-2-chlorophenylamino)-7-fluoro-3-methylbenzo[d]isoxazole-5-carboxylicacid (2-vinyloxyethoxy) amide (11a) (55.4 mg, 0.114 mmol) and aqueousHCl (1 M, 0.23 mL) in EtOH (3 mL) was stirred for 2 hours at roomtemperature. The pH of the reaction mixture was adjusted to 6-7 withaqueous NaOH (2 M). The reaction was diluted with EtOAc. The organiclayer was washed with water, dried over MgSO₄, filtered, andconcentrated in vacuo to give 50.2 mg (96% yield) of the desired product(12a). MS APCI (−) m/z 456, 458 (M+, Br, Cl pattern) detected. ¹H NMR(400 MHz, CD₃OD) δ 7.87 (s, 1H), 7.57 (d, 1H), 7.31 (dd, 1H), 6.74 (dd,1H), 4.01 (t, 2H), 3.74 (t, 2H), 2.60 (s, 3H): ¹⁹F NMR (376 MHz,CD₃OD)−140.85 (s).

Example 4

Synthesis ofN-[6-(4-bromo-2-chlorophenylamino)-7-fluoro-3-methylbenzo[d]isoxazole-5-carbonyl]-methanesulfonamide(13a)

A mixture of6-(4-bromo-2-chlorophenylamino)-7-fluoro-3-methylbenzo[d]isoxazole-5-carboxylicacid (9a) (41 mg, 0.102 mmol) and carbonyldiimidazole (23 mg, 0.140mmol) in THF (1 mL) was stirred at 50° C. in a sealed tube reactor. Thereaction mixture was cooled to room temperature and methanesulfonamide(17 mg, 0.179 mmol) was added followed by DBU (0.025 mL, 0.164 mmol).After stirring at 50° C. for 1 hour, the reaction mixture was cooled toroom temperature, and diluted with EtOAc. The organic layer was washedwith water, 1 N HCl, and brine. The organic layer was dried (MgSO₄) andconcentrated. Purification by flash column chromatography using theBiotage system (7% MeOH in CH₂Cl₂) provided 34 mg (65% yield) of thedesired product (13a). MS APCI (−) m/z 474, 476 (M+, Br, Cl pattern)detected. ¹H NMR (400 MHz, CD₃OD) δ 8.27 (s, 1H), 7.53 (s, 1H), 7.27 (d,1H), 6.73 (t, 1H), 3.11 (s, 3H), 2.55 (s, 3H): ¹⁹F NMR (376 MHz,CD₃OD)−141.84 (s, 1F).

Example 5

Synthesis of6-(2,4-dichlorophenylamino)-7-fluoro-3-methylbenzo[d]isoxazole-5-carboxylicacid (9b)

Step A: Preparation of6-(2,4-dichlorophenylamino)-7-fluoro-3-methylbenzo[d]isoxazole-5-carboxylicacid methyl ester (8b):6-(2-Chlorophenylamino)-7-fluoro-3-methylbenzo[d]isoxazole-5-carboxylicacid methyl ester (7) (129 mg, 0.384 mmol) and N-chlorosuccinimide (57mg, 0.421 mmol) were stirred in DMF (5 mL) for 16 hours. ConcentratedHCl (3 μL) was added and the reaction mixture stirred 2 hours. Thereaction mixture was diluted with EtOAc, and washed with water (2×). Theorganic layer was dried over MgSO₄, filtered, and concentrated.Purification by flash column chromatography using the Biotage system (5%EtOAc in hexanes) provided 73 mg (52%) the desired product (8b).

Step B: Preparation of6-(2,4-dichlorophenylamino)-7-fluoro-3-methylbenzo[d]isoxazole-5-carboxylicacid (9b): Compound 9b was prepared according to Step H of Example 1using6-(2,4-dichlorophenylamino)-7-fluoro-3-methylbenzo[d]isoxazole-5-carboxylicacid methyl ester (8b) to provide 68 mg (98% yield) of the desiredproduct (9b). MS APCI (−) m/z 353, 355 (M+, Br, Cl pattern) detected. ¹HNMR (400 MHz, DMSO-d₆) δ 9.58 (s, 1H), 8.34 (s, 1H), 7.65 (d, 1H), 7.31(dd, 1H), 7.04 (dd, 1H), 2.60 (s, 3H): ¹⁹F NMR (376 MHz, DMSO-d₆)−140.36(s).

Example 6

Synthesis of6-(2,4-dichlorophenylamino)-7-fluoro-3-methylbenzo[d]isoxazole-5-carboxylicacid (2-hydroxyethoxy)amide (12b)

The synthesis of compound 12b was carried out according to Steps A and Bof Example 3 using6-(2,4-dichlorophenylamino)-7-fluoro-3-methylbenzo[d]isoxazole-5-carboxylicacid (9b) as the starting material to provide 29 mg (38% yield for twosteps) of 12b. MS APCI (−) m/z 412, 414 (M+, Br, Cl pattern) detected.¹H NMR (400 MHz, CD₃OD) δ 7.87 (s, 1H), 7.45 (m, 1H), 7.19 (m, 1H), 6.80(m, 1H), 4.02 (t, 2H), 3.75 (t, 2H), 2.60 (s, 3H): ¹⁹F NMR (376 MHz,CD₃OD)−141.05 (s).

Example 7

Synthesis of3-amino-6-(4-bromo-2-chlorophenylamino)-7-fluorobenzo[d]isoxazole-5-carboxylicacid (19)

Step A: Preparation of2-(2-chlorophenylamino)-5-cyano-3,4-difluorobenzoic acid methyl ester(15): A mixture of 5-bromo-2-(2-chlorophenylamino)-3,4-difluorobenzoicacid methyl ester (14) (3.01 g, 7.99 mmol),1,1′-bis(diphenylphosphino)ferrocene (dppf) (93 mg, 0.162 mmol), Pd₂dba₃ (73 mg, 0.080 mmol) and Zn(CN)₂ (573 mg, 4.78 mmol) in1methyl-2-pyrrolidinone (NMP: 4.5 mL) was heated in a sealed tubereactor. After 20 hours, the reaction mixture was cooled to roomtemperature, quenched by the addition of 8 mL 4:1:4 (volume) mixture ofsaturated NH₄Cl, concentrated NH₄OH and water, and extracted with amixture of EtOAc/THF. The combined organic extracts were washed with4:1:4 (volume) mixture of saturated NH₄Cl, concentrated NH₄OH and water,and brine. The organic layer was dried (MgSO₄) and concentrated.Purification by flash column chromatography using the Biotage system(twice: 100% hexanes to 35% CH₂Cl₂ in hexanes, then 30% CH₂Cl₂ inhexanes) provided 1.33 g (52%) of the desired product (15).

Step B: Preparation of3-amino-6-(2-chlorophenylamino)-7-fluorobenzo[d]isoxazole-5-carboxylicacid methyl ester (17): t-BuOK (3.80 mL of a 1.0 M solution in THF) wasadded to a stirred solution of propan-2-one oxime (285 mg, 3.82 mmol) inTHF (5 mL) at room temperature. The reaction mixture was further dilutedwith THF (20 mL) and after 30 minutes cooled to 0° C. A solution of2-(2-chlorophenylamino)-5-cyano-3,4-difluorobenzoic acid methyl ester(15) (600 mg, 1.86 mmol) in THF (5 mL) was added. The reaction mixturewas slowly warmed to room temperature. After 90 minutes, the reactionmixture was quenched with saturated NH₄Cl and diluted with EtOAc. Theorganic layer was washed with saturated NH₄Cl and brine, dried (MgSO₄)and concentrated. The residue (16) was diluted with MeOH (10 mL) and asolution of 2 M HCl in diethyl ether (10 mL) was added. After 16 hours,the reaction mixture was diluted with EtOAc, washed with water,saturated NaHCO₃ and water. The organic layer was dried (MgSO₄) andconcentrated. Purification by flash column chromatography using theBiotage system (1.5% MeOH in CH₂Cl₂) provided 399 mg (64%) of thedesired product (17).

Step C: Preparation of3-amino-6-(4-bromo-2-chlorophenylamino)-7-fluorobenzo[d]isoxazole-5-carboxylicacid methyl ester (18): Compound 18 was prepared according to Step G ofExample 1 using compound 17 as the starting material.

Step D: Preparation of3-amino-6-(4-bromo-2-chlorophenylamino)-7-fluorobenzo[d]isoxazole-5-carboxylicacid (19): Compound 19 was prepared according to Step H of Example 1using3-amino-6-(4-bromo-2-chlorophenylamino)-7-fluorobenzo[d]isoxazole-5-carboxylicacid methyl ester (18) as the starting material to provide 188 mg (98%yield) of compound 19. MS APCI (−) m/z 398, 400 (M+, Br, Cl pattern)detected. ¹H NMR (400 MHz, DMSO-d₆) δ 9.47 (s, 1H), 8.49 (s, 1H), 7.73(m, 1H), 7.41 (dd, 1H), 6.92 (t, 1H), 6.76 (s, 2H): ¹⁹F NMR (376 MHz,DMSO-d₆)−141.48 (s).

Example 8

Synthesis of3-amino-6-(4-bromo-2-chloro-phenylamino)-7-fluorobenzo[d]isoxazole-5-carboxylicacid (2-hydroxyethoxy)-amide (21).

The synthesis of compound 21 was accomplished according to Steps A and Bof Example 3 using3-amino-6-(4-bromo-2-chlorophenylamino)-7-fluorobenzo[d]isoxazole-5-carboxylicacid (19) as the starting material to provide 16 mg (23% yield for twosteps) of compound 21. MS APCI (−) m/z 457, 459 (M+, Br, Cl pattern)detected. ¹H NMR (400 MHz, DMSO-d₆) δ 11.92 (s, 1H), 8.59 (s, 11H), 7.94(s, 11H), 7.69 (s, 1H), 7.36 (d, 11H), 6.75 (dd, 1H), 6.71 (s, 2H), 4.73(s, 1H), 3.87 (s, 2H), 3.59 (s, 2H): ¹⁹F NMR (376 MHz, DMSO-d₆)−140.64(s).

Example 9

Synthesis of7-(4-bromo-2-chlorophenylamino)-8-chloroimidazo[1,2-a]pyridine-6-carboxylicacid (30)

Step A: Preparation of 4,6-dichloronicotinic acid ethyl ester (22):POCl₃ (100 mL, 1092 mmol) was added to 4,6-dihydroxynicotinic acid ethylester (J. Heterocyclic Chem. 1983, 20, 1363) (20.0 g, 109 mmol). Theresulting suspension was cooled to 0° C. and triethylamine (15.2 mL, 109mmol) was added dropwise at such a rate as to maintain the internalreaction mixture temperature below 25° C. Upon completion of addition,the reaction mixture was warmed to room temperature and then to 80° C.After 4 hours, the reaction mixture was cooled to room temperature andstirred for 16 hours. The reaction mixture was carefully poured onto 2 Lcrushed ice. The mixture was extracted with EtOAc and diethyl ether. Thecombined organic extracts were washed with brine, dried (Na₂SO₄) andconcentrated. The dark brown liquid was purified by passing through aplug of silica gel (CH₂Cl₂) to give the desired product (22) as a lowmelting yellow solid (18.7 g, 78%).

Step B: Preparation of 4.6-dichloronicotinic acid (23): Sodium hydroxide(40 mL, 6.25 M solution) was added to a stirred solution of4,6-dichloronicotinic acid ethyl ester (22) (25.95 g, 118 mmol) in 4:1:1THF/MeOH/water (600 mL). After 30 minutes, the reaction mixture wasacidified to pH 2 with concentrated HCl, diluted with 1:1 EtOAc/Et₂O andwashed with water and brine. The organic layer was dried (Na₂SO₄) andconcentrated. The resulting off-white solid was twice concentrated fromtoluene to give the desired product (23) as a white solid (21.73 g,96%).

Step C: Preparation of 4-(4-bromo-2-chlorophenylamino)-6-chloronicotinicacid hydrochloride salt (24): LiHMDS (261 mL of a 1 M solution inhexanes) was added dropwise over 30 minutes to a solution of4-bromo-2-chlorophenylamine (35.0 g, 172 mmol) in THF (80 mL) at −78° C.After 1 hour, 4,6-dichloronicotinic acid (23) (15.7 g, 81.7 mmol) wasadded dropwise over 30 minutes. The reaction mixture was slowly warmedto room temperature and stirred 16 hours. The reaction mixture wasquenched with water, diluted with EtOAc and acidified with 1 M HCl. Theresulting precipitate was isolated by filtration and washed with EtOAc.The solids were twice concentrated from toluene, triturated with CH₂Cl₂and collected by filtration. The solids were further concentrated fromtoluene (3×) followed by drying in vacuo to give the desired product(24) containing a small amount of water (36.0 g).

Step D: Preparation of4-(4-bromo-2-chlorophenylamino)-5,6-dichloronicotinic acid (25):N-Chlorosuccinimide was (13.0 g, 99.0 mmol) added to a suspension of4-(4-bromo-2-chlorophenylamino)-6-chloronicotinic acid (24) (32.54 g,89.9 mmol) in DMF (500 mL). The suspension was allowed to stir at roomtemperature overnight. The reaction mixture was diluted with saturatedsodium bisulfite (200 mL) and water (1L) resulting in formation of athick white precipitate which was isolated by filtration and washed withwater. The solids were dissolved into THF. Two volumes of diethyl etherwere added and the organic solution washed with brine, dried over NaSO₄,filtered, and concentrated in vacuo to provide an orange solid. Thesolid was triturated with diethyl ether to provide the desired productas an off-white solid (25) (13.34 g, 37%). MS (APCI−) m/z 393, 395, 397(M-; Cl, Br pattern) detected.

Alternatively, 4-(4-bromo-2-chlorophenylamino)-5,6-dichloronicotinicacid (25) can be synthesized by the route and procedure described below.

N-Chlorosuccinimide (56.5 g, 423 mmol) was added portionwise to asuspension of 4,6-dihydroxynicotinic acid ethyl ester (70.5 g, 385 mmol)in DMF (705 mL). Concentrated HCl (3.20 mL, 38.5 mmol) was added. Afterstirring for 2.5 hours, the product was precipitated with water andNa₂S₂O₃ (70 mL). The slurry was acidified to pH 3 with 2 M HCl (30 mL).5-Chloro-4,6-dihydroxynicotinic acid ethyl ester, the desired productwas isolated as a pale yellow solid (75.7 g, 90%) by filtration. MS ESI(+) m/z 218, 220 (M+, Cl pattern) detected.

5-Chloro-4,6-dihydroxynicotinic acid ethyl ester (8.05 g, 37 mmol) wassuspended in phosphorous oxychloride (30 mL, 296 mmol). The mixture wascooled to 0° C. and triethylamine (5.16 mL, 37.0 mmol) was added. Thereaction was heated to 60° C. for three hours. The solution was cooledto room temperature, poured onto ice, stirred for 15 minutes andextracted with ethyl acetate (2×) and diethyl ether (1×). The combinedorganic extracts were washed with brine (3×), dried over Na₂SO₄ andconcentrated to a brown liquid. The crude product was passed through aplug of silica gel eluting with dichloromethane.4,5,6-trichloronicotinic acid ethyl ester, the desired product wasobtained as a yellow liquid (7.76 g, 82%).

Sodium hydroxide (1.0 M solution, 61.0 mL, 61.0 mmol) was added to asolution of 4,5,6-trichloronicotinic acid ethyl ester (7.76 g, 30.5mmol) in 4:1 THF/MeOH (150 mL). After stirring for 30 minutes, thereaction was acidified to pH 1 by addition of concentrated HCl, dilutedwith ethyl acetate, washed with water (3×) and brine (2×), dried overNa₂SO₄ and concentrated to provide the desired product,4,5,6-trichloro-nicotinic acid, as an off white solid (6.77 g, 98%).

LiHMDS (1.0 M solution in hexanes, 53.0 mL, 53.0 mmol) was addeddropwise to a stirred solution of 4-bromo-2-chlorophenylamine (7.10 g,35.0 mmol) in THF (15 mL) cooled to −78° C. After one hour,4,5,6-trichloronicotinic acid (3.73 g, 16.5 mmol) was added dropwise asa solution in THF (12 mL). The reaction was allowed to warm to roomtemperature slowly while stirring overnight. The suspension was dilutedwith 1 M HCl and ethyl acetate, and the aqueous phase was extracted withethyl acetate (3×). The combined organic phases were dried over Na₂SO₄and concentrated to a tan solid. The solid was triturated with diethylether overnight and the solids were isolated by filtration to providethe desired product (25) as a tan-colored solid (4.85 g, 75%). MS APCI(−) m/z 395, 397 (M-, Cl, Br pattern) detected.

Step E: Preparation of4-(4-bromo-2-chlorophenylamino)-5,6-dichloronicotinic acid methyl ester(26): Trimethylsilyldiazomethane (2.0 M solution in hexanes, 37 mL, 74mmol) was added slowly to a suspension of4-(4-bromo-2-chloro-phenylamino)-5,6-dichloronicotinic acid (25) (14.67g, 37 mmol). After the addition was complete the resulting slurry wasdiluted with hexanes (600 mL) and the solids isolated by filtrationwashing with hexanes. The desired product was isolated as an off-whitesolid (10.06 g). The hexanes washes were concentrated and the solidspassed through a plug of silica gel eluting with dichloromethane.Concentration of the product-containing fractions provided an additional3.83 g desired product (26) for a total of 13.89 g (91%). MS (APCI+) m/z409, 411, 413 (M+; Cl, Br pattern) detected.

Step F: Preparation of6-azido-4-(4-bromo-2-chlorophenylamino)-5-chloronicotinic acid methylester (27): Sodium azide (4.4 g, 68 mmol) was added to a suspension of4-(4-bromo-2-chlorophenylamino)-5,6-dichloronicotinic acid methyl ester(26) (13.89 g, 33.8 mmol) in DMF (200 mL) and the mixture allowed tostir at room temperature overnight. The solution was diluted with water(600 mL) and the resulting white precipitate was collected by filtrationand washed with water. The solids were dissolved into THF. Two volumesof diethyl ether were added and the organic solution washed with brine,dried over NaSO₄, filtered, and concentrated in vacuo to the desiredproduct (27) as a light yellow solid (12.94 g, 92%).

Step G: Preparation of6-amino-4-(4-bromo-2-chlorophenylamino)-5-chloro-nicotinic acid methylester (28): Zinc powder (10 g, 155 mmol) was added portionwise to asuspension of 6-azido-4-(4-bromo-2-chlorophenylamino)-5-chloronicotinicacid methyl ester (27) (12.94 g, 31 mmol) in 3:1 dichloromethane/aceticacid (300 mL). After fifteen minutes the reaction mixture was pouredinto 700 mL ethyl acetate, washed with water, saturated sodiumbicarbonate and brine. The organic solution was dried over NaSO₄,filtered, and concentrated in vacuo to provide the desired product (28)as an off-white solid (11.85 g, 98%). MS (APCI+) m/z 390, 392, 394 (M+;Cl, Br pattern) detected.

Step H: Preparation of7-(4-bromo-2-chlorophenylamino)-8-chloroimidazo[1,2-a]pyridine-6-carboxylicacid methyl ester (29): Chloroacetaldehyde (50% aqueous solution, 0.70mL, 5.7 mmol) was added to a suspension of6-amino-4-(4-bromo-2-chlorophenylamino)-5-chloronicotinic acid methylester (28) in DMF (7 mL) contained in a sealed tube. The reactionmixture was heated at 80° C. for four hours and then allowed to cool toroom temperature and stir overnight. The dark brown solution was dilutedwith water (70 mL) the resulting light brown precipitate was collectedby filtration and washed with water. The solids were dissolved into THF.Two volumes of ethyl acetate were added and the organic solution washedwith brine, dried over NaSO₄, filtered, and concentrated in vacuo toprovide a brown solid. The aqueous filtrate was extracted with ethylacetate and the organic extracts were dried over NaSO₄, filtered, andconcentrated in vacuo. This material was combined with the previouslyisolated brown solid and the combined material subjected to columnchromatography (dichloromethane, followed by 20:1dichloromethane/methanol). The desired product (29) was isolated as alight yellow solid (0.752 g, 64%). MS (APCI+) m/z 414, 416, 418 (M+; Cl,Br pattern) detected.

Step I: Preparation of7-(4-bromo-2-chlorophenylamino)-8-chloroimidazo[1,2-a]pyridine-6-carboxylicacid (30): Sodium hydroxide (1.0 M aqueous solution, 14.6 mL, 14.6 mmol)was added to a solution of7-(4-bromo-2-chlorophenylamino)-8-chloroimidazo[1,2-a]pyridine-6-carboxylicacid methyl ester (29) in methanol (30 mL) and the solution allowed tostir at room temperature overnight. Methanol was removed by rotaryevaporation and the solution diluted with water and acidified to pH 2 byaddition of 1.0 M HCl. The aqueous suspension was extracted with 4:1ethyl acetate/THF. The organic extracts were washed with brine, driedover NaSO₄, filtered, and concentrated in vacuo to provide the desiredproduct as a light orange solid (30). MS (APCI+) m/z 400, 402, 404 (M+:Cl, Br pattern) detected. ¹H NMR (400 MHz, methanol-d₄) δ 9.01 (s, 1H),7.83 (s, 1H), 7.51 (s, 2H), 7.25 (d, 1H), 6.60 (d, 1H).

The following compounds were synthesized in a similar manner asdescribed in Example 9.

8-Chloro-7-(2,4-dichlorophenylamino)-imidazo[1,2-a]pyridine-6-carboxylicacid

MS APCI (+) m/z 356, 358 (M+, Cl pattern) detected. ¹H NMR (400 MHz,CD₃OD) δ 9.16 (s, 1H), 8.00 (d, 1H), 7.72 (d, 1H), 7.51 (d, 1H), 7.25(dd, 1H), 7.02 (d, 1H).

8-Chloro-7-(2-fluoro-4-iodophenylamino)-imidazo[1,2-a]pyridine-6-carboxylicacid

MS APCI (+) m/z 432, 434 (M+, Cl pattern) detected. ¹H NMR (400 MHz,DMSO-d₆) δ 9.31 (s, 1H), 8.11 (s, 1H), 7.71 (s, 1H), 7.61 (d, 1H), 7.37(d, 1H), 6.62 (t, 2H).

8-Chloro-7-(2,4-difluorophenylamino)-imidazo[1,2-a]pyridine-6-carboxylicacid

MS APCI (+) m/z 324, 326 (M+, Cl pattern) detected. ¹H NMR (400 MHz,DMSO-d₆) δ 9.27 (s, 1H), 8.06 (s, 1H), 7.65 (s, 1H), 7.29 (t, 1H), 6.96(m, 2H). ¹⁹F (376 MHz, DMSO-d₆)−118.9 (s, 1F), −124.8 (s, 1F).

7-(4-Bromo-2-methyl-phenylamino)-8-chloroimidazo[11,2-a]pyridine-6-carboxylicacid

MS APCI (+) m/z 380, 382 (M+, Cl, Br pattern) detected. ¹H NMR (400 MHz,DMSO-d₆) δ 9.31 (s, 11H), 8.09 (s, 11H), 7.69 (s, 11H), 7.42 (s, 11H),7.24 (d, 11H), 6.64 (d, 1H), 2.28 (s, 3H).

Example 10

1-[7-(4-Bromo-2-chlorophenylamino)-8-chloroimidazo[1,2-a]pyridin-6-yl]-ethanone

This compound is prepared from compound 29, the product of Example 9,Step H. Tebbe reagent(μ-chloro-μ-methylene[bis(cyclopentadienyl)titanium]dimethyl-aluminum, 1M solution in toluene, 0.12 mL, 0.12 mmol) was added to a solution of7-(4-bromo-2-chloro-phenylamino)-8-chloro-imidazo[1,2-a]pyridine-6-carboxylicacid methyl ester (29) (25 mg, 0.061 mmol) in THF (1 mL) cooled to 0° C.The reaction mixture was warmed to room temperature and stirred for 1.5hours. HCl (10% aqueous solution, 1 mL) was added and the mixturestirred for 16 hours. The reaction was diluted with ethyl acetate,washed with saturated aqueous sodium carbonate, dried over MgSO₄, andconcentrated. The crude material was purified by flash columnchromoatography (dichloromethane to 100:1 dichloromethane/methanol) toprovide the desired product (6.8 mg, 28%). MS APCI (−) m/z 396, 398, 400(M-, Cl, Br pattern) detected. ¹H NMR (400 MHz, CDCl₃) δ 9.05 (br s,1H), 8.81 (s, 1H), 7.71 (s, 2H), 7.55 (d, 1H), 7.22 (dd, 1H), 6.55 (d,1H), 2.67 (s, 3H).

Example 11 Preparation of3-bromo-7-(4-bromo-2-chlorophenylamino)-8-chloro-imidazo[1,2-a]pyridine-6-carboxylicacid (2-hydroxyethoxy)-amide

Step A: Preparation of3-bromo-7-(4-bromo-2-chlorophenylamino)-8-chloroimidazo[1,2-a]pyridine-6-carboxylicacid methyl ester. N-bromosuccinimide (14 mg, 0.080 mmol) was added to asolution of7-(4-bromo-2-chlorophenylamino)-8chloroimidazo[1,2-a]pyridine-6-carboxylicacid methyl ester (30 mg, 0.072 mmol) in chloroform (1.0 mL). Afterstirring for five hours the reaction was diluted with ethyl acetate,washed with NaHSO₃, brine, dried over Na₂SO₄, and concentrated to ayellow solid (33 mg, 92%). MS APCI (+) m/z 494, 496, 498 (M+, Cl, Brpattern) detected. ¹H NMR (400 MHz, CDCl₃) δ 8.82 (s, 1H), 8.65 (s, 1H),7.67 (s, 1H), 7.55 (dd, 1H), 7.22 (dd, 1H), 6.53 (d, 1H), 3.98 (s, 3H).

Step B: Preparation of3-bromo-7-(4-bromo-2-chlorophenylamino)-8-chloroimidazo[1,2-a]pyridine-6-carboxylicacid (2-hydroxyethoxy)-amide. The synthesis of the title compound wascarried out according to Step H of Example 1 and Steps A and B ofExample 3 using3-bromo-7-(4-bromo-2-chlorophenylamino)-8-chloroimidazo[1,2-a]pyridine-6-carboxylicacid methyl ester as the starting material to provide 20 mg of desiredproduct as a light yellow solid. MS APCI (+) m/z 539, 541, 543 (M+, Cl,Br pattern) detected.

Example 12 Preparation of7-(4-romo-2-chlorophenylamino)-8-chloro-3-methylimidazo[1,2-a]pyridine-6-carboxylicacid cyclopropylmethoxyamide

Step A: Preparation of4-(4-bromo-2-chloroiphenylamino)-5,6-dichloronicotinic acid tert-butylester. 2-tert-Butyl-1,3-diisopropyl-isourea (10.6 g, 53 mmol) was addedto a solution of 4-(4-bromo-2-chlorophenylamino)-5,6-dichloro-nicotinicacid (4.21 g, 10.6 mmol) in THF (200 mL). The reaction was heated toreflux. After 30 minutes the reaction was cooled to room temperature anddiluted with EtOAc. The organic layer was washed with saturated K₂CO₃(2×), brine, dried over Na₂SO₄ and concentrated. The yellow solid wastriturated with dichloromethane and white solids were removed byfiltration. The filtrate was concentrated in vacuo to yield the desiredproduct as a yellow solid (5.53 g, 97%). MS APCI (−) m/z 451, 453 (M-,Cl, Br pattern) detected.

Step B: Preparation of4-(4-bromo-2-chlorophenylamino)-5-chloro-6-(2-hydroxypropylamino)-nicotinicacid tert-butyl ester. I-Amino-propan-2-ol (2.44 g, 30.3 mmol) andtriethylamine (0.42 mL, 3.03 mmol) were added to a solution of4-(4-bromo-2-chlorophenylamino)-5,6-dichloronicotinic acid tert-butylester (1.37 g, 3.03 mmol) in acetonitrile (30 mL). The reaction washeated to reflux. After 23 hours the reaction was cooled to roomtemperature and diluted with EtOAc, washed with saturated NaHCO₃, water,brine, dried over Na₂SO₄ and concentrated to a white solid. Purificationby flash column chromatography (20:1 dichloromethane/methanol) providedthe desired product as a white solid (1.11 g, 75%). MS APCI (+) m/z 492,494 (M+, Cl, Br pattern) detected.

Step C: Preparation of4-(4-bromo-2-chlorophenylamino)-5-chloro-6-(2-oxopropylamino)-nicotinicacid tert-butyl ester. To a solution of4-(4-bromo-2-chlorophenylamino)-5-chloro-6-(2-hydroxypropylamino)-nicotinicacid tert-butyl ester (0.28 g, 0.57 mmol) in acetonitrile (1.1 mL) wasadded 4A molecular sieves and N-methylmorpholine (0.10 g, 0.85 mmol).The mixture was cooled to 0° C. and tetrapropylammonium perruthenate(0.030 g, 0.085 mmol) was added. After stirring for one hour, thereaction was filtered through a plug of silica gel, washing with EtOAc.The filtrate was concentrated. Purification by flash columnchromatography (20:1 hexanes/ethyl acetate) provided the desired productas a white solid (86 mg, 31%). MS APCI (+) m/z 490, 492 (M+, Cl, Brpattern) detected.

Step D: Preparation of7-(4-bromo-2-chlorophenylamino)-8-chloro-3-methylimidazo[1,2-a]pyridine-6-carboxylicacid.4-(4-Bromo-2-chlorophenylamino)-5-chloro-6-(2-oxopropylamino)-nicotinicacid tert-butyl ester (0.075 g, 0.15 mmol) was dissolved intoconcentrated H₂SO₄ (0.50 mL). After ten minutes, ice and water were andthe mixture was stirred for ten minutes. The mixture was diluted withEtOAc, neutralized with 1 M NaOH, washed with brine, dried over Na₂SO₄and concentrated to provide the desired product as an off-white solid.MS APCI (+) m/z 416, 418 (M+, Cl, Br pattern) detected.

Step E: Preparation of7-(4-bromo-2-chlorophenylamino)-8-chloro-3-methylimidazo[1,2-a]pyridine-6-carboxylicacid cyclopropylmethoxy-amide. The synthesis of the title compound wascarried out according to Example 2 using7-(4-bromo-2-chlorophenylamino)-8-chloro-3-methylimidazo[1,2-a]pyridine-6-carboxylicacid as the starting material to provide 8 mg (18%) of desired productas a yellow solid. MS APCI (+) m/z 485, 487 (M+, Cl, Br pattern)detected. ¹H NMR (400 MHz, CDCl₃) δ 8.68 (s, 1H), 7.72 (m, 1H), 7.54 (m,1H), 7.20 (dd, 1H), 6.42 (d, 1H), 3.58 (d, 2H), 2.57 (s, 3H), 0.64 (m,1H), 0.57 (m, 2H), 0.23 (m, 2H).

Example 13

7-(4-bromo-2-chlorophenylamino)-8-chloroimidazo[1,2-a]pyridine-6-carboxylicacid cyclopropylmethoxyamide (31)

Compound 31 was prepared according to the method of Example 2, using7-(4-bromo-2-chlorophenylamino)-8-chloroimidazo[1,2-a]pyridine-6-carboxylicacid (30) as the starting material to provide 4.1 g (53% yield) ofcompound 31. MS (APCI−) m/z 467, 469, 471 (M-: Cl, Br pattern) detected.¹H NMR (400 MHz, CD₃OD) δ 8.76 (s, 1H), 7.95 (d, 1H), 7.64 (d, 1H), 7.56(d, 1H), 7.26 (dd, 1H), 6.56 (d, 1H), 3.57 (d, 2H), 1.10 (m, 1H), 0.54(m, 2H), 0.24 (m, 2H).

The following compounds were synthesized in a similar manner asdescribed in Example 9 using the appropriate aniline in Step C.

7-(4-Bromo-2-fluorophenylamino)-8-chloroimidazo[1,2-a]pyridine-6-carboxylicacid cyclopropylmethoxyamide

MS ESI (+) m/z 453, 455, 457 (M+, Cl, Br pattern) detected. ¹H NMR (400MHz, CD₃OD) δ 8.70 (s, 1H), 7.95 (s, 1H), 7.67 (s, 1H), 7.34 (m, 1H),7.20 (m, 1H), 6.79 (m, 1H), 3.49 (m, 2H), 1.08 (m, 1H), 0.55 (m, 2H),0.26 (m, 2H). ¹⁹F NMR (376 MHz, CD₃OD) δ−127.4.

8-chloro-7-(2-fluorophenylamino)-imidazo[1,2-a]pyridine-6-carboxylicacid cyclopropylmethoxyamide

MS ESI (+) m/z 375, 377 (M+, Cl pattern) detected. ¹H NMR (400 MHz,CD₃OD) δ 8.70 (s, 1H), 7.91 (s, 1H), 7.60 (s, 1H), 7.09 (m, 1H), 7.00(m, 1H), 6.95 (m, 1H), 6.77 (m, 1H), 3.47 (d, 2H), 1.05 (m, 1H), 0.51(m, 2H), 0.22 (m, 2H). ¹⁹F NMR (376 MHz, CD₃OD) δ −132.1.

7-(4-bromo-2-fluorophenylamino)-8-chloroimidazo[1,2-a]pyridine-6-carboxylicacid (2-hydroxyethoxy)-amide

MS ESI (+) m/z 443, 445, 447 (M+, Cl, Br pattern) detected. ¹H NMR (400MHz, CD₃OD) δ 8.74 (s, 1H), 7.91 (s, 1H), 7.61 (s, 1H), 7.32 (m, 1H),7.16 (m, 1H), 6.68 (m, 1H), 3.84 (t, 2H), 3.66 (t, 2H). ¹⁹F NMR (376MHz, CD₃OD) δ−128.9.

8-Chloro-7-(2-fluorophenylamino)-imidazo[1,2-a]pyridine-6-carboxylicacid (2-hydroxyethoxy)-amide

MS ESI (+) m/z 365, 367 (M+, Cl pattern) detected. ¹H NMR (400 MHz,CD₃OD) δ 8.73 (s, 1H), 7.89 (s, 1H), 7.59 (s, 1H), 7.10 (m, 1H), 7.00(m, 1H), 6.94 (m, 1H), 6.77 (m, 1H), 3.78 (t, 2H), 3.62 (t, 2H). ¹⁹F NMR(376 MHz, CD₃OD) δ−131.9.

8-chloro-7-(2,4-dichlorophenylamino)-imidazo[1,2-a]pyridine-6-carboxylicacid (2-hydroxyethoxy) amide

MS APCI (−) m/z 413, 415, 417 (M-, Cl pattern) detected. ¹H NMR (400MHz, CD₃OD) δ 8.78 (s, 1H), 7.90 (s, 1H), 7.87 (s, 1H), 7.10 (dd, 1H),6.61 (d, 1H), 4.0 (m, 2H), 3.72 (m, 2H).

7-(4-Bromo-2-fluorophenylamino)-8-chloroimidazo[1,2-a]pyridine-6-carboxylicacid

MS ESI (+) m/z 384, 386, 388 (M+, Cl, Br pattern) detected. ¹H NMR (400MHz, DMSO-d₆) δ 9.29 (s, 1H), 8.10 (s, 1H), 7.68 (s, 1H), 7.53 (m, 1H),7.23 (m, 1H), 6.75 (m, 1H). ¹⁹F NMR (376 MHz, DMSO-d₆) δ −127.9.

8-Chloro-7-(2-fluorophenylamino)-imidazo[1,2-a]pyridine-6-carboxylicacid

MS ESI (+) m/z 306, 308 (M+, Cl pattern) detected. ¹H NMR (400 MHz,DMSO-d₆) δ 9.30 (s, 1H), 8.09 (s, 1H), 7.67 (s, 1H), 7.22 (dd, 1H), 7.06(dd, 1H), 6.98 (m, 1H), 6.84 (m, 1H). ¹⁹F NMR (376 MHz, DMSO-d₆) δ−130.5.

8-Chloro-7-(4-chloro-2-fluorophenylamino)-imidazo[1,2-a]pyridine-6-carboxylicacid

MS ESI (+) m/z 340, 342 (M+, Cl pattern) detected. ¹H NMR (400 MHz,DMSO-d₆) δ 9.29 (s, 1H), 8.10 (s, 1H). 7.68 (s, 1H), 7.43 (m, 1H), 7.12(m, 1H), 6.83 (m, 1H). ¹⁹F NMR (376 MHz, DMSO-d₆) δ −127.8.

8-Chloro-7-(4-chloro-2-fluorophenylamino)-imidazo[1,2-a]pyridine-6-carboxylicacid cyclopropylmethoxyamide

MS ESI (+) m/z 409, 411 (M+, Cl pattern) detected. ¹H NMR (400 MHz,CD₃OD) δ 9.97 (br s, 1H), 8.82 (s, 1H). 7.73 (s, 1H), 7.71 (s, 1H), 7.18(m, 1H), 6.97 (m, 1H), 6.56 (m, 1H), 6.47 (br s, 1H), 3.60 (m, 2H), 1.00(m, 1H), 0.56 (m, 2H), 0.24 (m, 2H). ¹⁹F NMR (376 MHz, CD₃OD) δ−128.7.

Example 14

Synthesis of7-(4-bromo-2-chlorophenylamino)-8-chloroimidazo[1,2-a]pyridine-6-carboxylicacid (2-hydroxyethoxy)-amide (33a)

Compound 33a was prepared according to Steps A and B of Example 3 using7-(4-bromo-2-chlorophenylamino)-8-chloroimidazo[1,2-a]pyridine-6-carboxylicacid (30) to provide 44 mg (40% yield for two steps) of the desiredproduct. MS (APCI+) m/z 459, 461, 463 (M+: Cl, Br pattern) detected. ¹HNMR (400 MHz, methanol-d₄) δ 8.90 (s, 1H), 8.08 (s, 1H), 7.93 (s, 1H),7.69 (s, 1H), 7.45, (d, 1H), 7.06 (m, 1H), 3.86 (br s, 2H), 3.72 (br s,2H).

Example 15

Synthesis of7-(4-bromo-2-chlorophenylamino)-8-chloro-3-(4-methylpiperazin-1-yl)-imidazo[1,2-a]pyridine-6-carboxylicacid cyclopropylmethoxyamide (36)

Step A:7-(4-Bromo-2-chlorolphenylamino)-8-chloro-3-(4-methylpiperazin-1-yl)-imidazo[1,2-a]pyridine-6-carboxylicacid methyl ester (34): Preparation was accomplished by modification ofthe procedure of Katritzky et al. (J. Org. Chem., 2003, 68, 4935-4937:J. Org. Chem., 1990, 55, 3209-3213). Bis(benzotriazazole) adduct (formedwith 1-methylpiperazine) (106 mg, 0.230 mmol) was added to a suspensionof 6-amino-4-(4-bromo-2-chlorophenylamino)-5-chloronicotinic acid methylester (28) (30 mg, 0.076 mmol) in dichloroethylene (1 mL) followed bythe addition of ZnBr₂ (52 mg, 0.230 mmol). The reaction mixture wasstirred at reflux for 10 hours and then at room temperature for 16hours. The reaction mixture was diluted with CH₂Cl₂ and filtered. Thefiltrate was washed with water. The aqueous layer was extracted withCH₂Cl₂. The combined organic extracts were washed with brine, dried(Na₂SO₄) and concentrated. Purification by flash column chromatographyusing the Biotage system (60:1 CH₂Cl₂:MeOH) provided the desired product(34) as a yellow solid (31 mg, 79%).

Step B:7-(4-Bromo-2-chlorophenylamino)-8-chloro-3-(4-methylpiperazin-1-yl)-imidazo[1,2-a]pyridine-6-carboxylicacid cyclopropylmethoxyamide (36): Sodium hydroxide (59 μL, 1 Msolution) was added to a suspension of7-(4-bromo-2-chlorophenylamino)-8-chloro-3-(4-methylpiperazin-1-yl)-imidazo[1,2-a]pyridine-6-carboxylicacid methyl ester (34) in MeOH (1 mL). After stirring 18 hours, thereaction mixture was concentrated to dryness. The residue (35) wasdiluted with toluene and concentrated (repeated), and 31 mg of therecovered yellow residue (35) was carried forward without purification.The residue (35) was suspended in CH₂Cl₂ (1 mL), cooled to 0° C. andoxalyl chloride (150 μL of a 2 M solution in CH₂Cl₂) was added. One dropof DMF was added and the reaction mixture warmed to room temperature.After 10 minutes, concentration of the mixture was followed byconcentrating from toluene twice and then drying in vacuo. The resultingyellow solid was suspended in CH₂Cl₂ (1 mL), cooled to 0° C. andcyclopropylmethylhydroxylamine (16 mg, 0.180 mmol) was added. After thereaction mixture was warmed to room temperature and stirred for 16hours, it was diluted with EtOAc. The organic layer was washed withsaturated NaHCO₃ and brine, dried (Na₂SO₄) and concentrated.Purification by flash column chromatography using the Biotage system(15:1 CH₂Cl₂/MeOH) provided the desired product (36) as a pale yellowsolid (12 mg, 37%). MS ESI (+) m/z 567, 569, 571 (M+, Cl, Br pattern)detected. ¹H NMR (400 MHz, CD₃OD) δ 8.35 (s, 1H), 7.55 (d, 1H), 7.38 (s,1H), 7.25 (dd, 1H), 6.54 (d, 1H), 3.59 (d, 2H), 3.17 (t, 4H), 2.74 (m,4H), 2.43 (s, 3H), 1.09 (m, 1H), 0.54 (m, 2H), 0.24 (m, 2H).

Example 16

Synthesis of7-(4-bromo-2-chlorophenylamino)-8-chloro-3-morpholin-4-yl-imidazo[1,2-a]-pyridine-6-carboxylicacid cyclopropylmethoxyamide (37)

Compound 37 was prepared according to Steps A and B of Example 15 using6-amino-4-(4-bromo-2-chlorophenylamino)-5-chloronicotinic acid methylester (28) and the bis(benzotriazazole) adduct (formed with morpholine)to provide 2 mg (8% yield for two steps) of the desired product (37). MSESI (+) m/z 554, 556, 558 (M+, Cl, Br pattern) detected. ¹H NMR (400MHz, CD₃OD) δ 8.41 (s, 1H), 7.55 (d, 1H), 7.38 (s, 1H), 7.26 (dd, 1H),6.54 (d, 1H), 3.91 (t, 4H), 3.59 (d, 2H), 3.11 (t, 4H), 1.08 (m, 1H),0.54 (m, 2H), 0.24 (m, 2H).

Example 17

Synthesis of7-(4-bromo-2-chlorophenylamino)-8-chloro-3-dimethylaminoimidazo[1,2-a]pyridine-6-carboxylicacid cyclopropylmethoxyamide (38)

Compound 38 was prepared according to Steps A and B of Example 15 using6-amino-4-(4-bromo-2-chlorophenylamino)-5-chloronicotinic acid methylester (28) and the bis(benzotriazazole) adduct (formed withdimethylamine) providing 16 mg (37% yield for two steps) of the desiredproduct (38). MS ESI (+) m/z 512, 514, 516 (M+, Cl, Br pattern)detected. ¹H NMR (400 MHz, CD₃OD) δ 8.37 (s, 1H), 7.54 (d, 1H), 7.30 (s,1H), 7.24 (dd, 1H), 6.52 (d, 1H), 3.59 (d, 2H), 2.86 (s, 6H), 1.07 (m,1H), 0.53 (m, 2H), 0.23 (m, 2H).

Example 18

Synthesis of7-(4-bromo-2-chlorophenylamino)-8-chloro-3-piperidin-1-ylmethylimidazo[1,2-a]pyridine-6-carboxylicacid (2-hydroxyethoxy) amide (39)

Compound 39 was prepared by a modification of the procedure of T.Kercher et al. (manuscript in preparation). Piperidine (4 μL, 0.043mmol) and 37% aqueous formaldehyde (5 μL, 0.065 mmol) were dissolved in6:1 MeCN:water (0.5 ml), and stirred 30 minutes.7-(4-bromo-2-chlorophenylamino)-8-chloroimidazo[1,2-a]pyridine-6-carboxylicacid (2-hydroxyethoxy)amide (33a) (10 mg, 0.022 mmol) was added followedby scandium triflate (1 mg, 0.002 mmol). After stirring 16 hours,additional scandium triflate (1 mg), piperidine (3.8 μL) and aqueousformaldehyde (3.8 μL) were added. After about 60 hours, the reactionmixture was diluted with EtOAc and washed with water, 10% K₂CO₃, andbrine. The organic layer was dried (Na₂SO₄) and concentrated.Purification by flash column chromatography using the Biotage system(40:1 CH₂Cl₂/MeOH to 20:1 CH₂Cl₂:MeOH to 9:1 CH₂Cl₂/MeOH) provided thedesired product (39) as a white solid (6 mg, 50%). MS APCI (+) m/z 556,558, 560 (M+, Cl, Br pattern) detected. ¹H NMR (400 MHz, CD₃OD) δ 8.83(s, 1H), 7.56 (s, 1H), 7.54 (s, 1H), 7.27 (dd, 1H), 6.56 (d, 1H), 3.91(m, 4H), 3.70 (m, 2H), 2.51 (broad s, 4H), 1.60 (broad s, 4H), 1.50(broad s, 2H).

The following compounds were synthesized in a similar manner asdescribed in Example 18.

7-(4-Bromo-2-chlorophenylamino)-8-chloro-3-morpholin-4-ylmethyl-imidazo[1,2-a]pyridine-6-carboxylicacid cyclopropylmethoxyamide

The reaction scheme for the synthesis of7-(4-bromo-2-chlorophenylamino)-8-chloro-3-morpholin-4-ylmethylimidazo[1,2-a]pyridine-6-carboxylicacid cyclopropylmethoxyamide is similar to that described in Example 18using7-(4-bromo-2-chlorophenylamino)-8-chloroimidazo[1,2-a]pyridine-6-carboxylicacid (2-hydroxyethoxy)amide (33a) and morpholine to provide the desiredproduct. MS APCI (+) m/z 568, 570, 572 (M+, Cl, Br pattern) detected; ¹HNMR (400 MHz, CDCl₃) δ 8.76 (s, 1H), 8.04 (s, 1H), 7.56 (d, 1H), 7.21(dd, 1H), 6.68 (d, 1H), 4.51 (s, 2H), 4.00 (m, 4H), 3.78 (d, 2H), 1.68(m, 1H), 0.56 (m, 2H), 0.26 (m, 2H).

7-(4-Bromo-2-chlorophenylamino)-8-chloro-3-dimethylaminomethylimidazo[1,2-a]pyridine-6-carboxylicacid cyclopropylmethoxyamide

The reaction scheme for the synthesis of7-(4-bromo-2-chlorophenylamino)-8-chloro-3-dimethylaminomethylimidazo[1,2-a]pyridine-6-carboxylicacid cyclopropylmethoxyamide was similar to that described in Example18, using7-(4-bromo-2-chlorophenylamino)-8-chloroimidazo[1,2-a]pyridine-6-carboxylicacid (2-hydroxyethoxy)amide (33a) and dimethylamine to provide thedesired product. MS APCI (+) m/z 528, 530, 532 (M+, Cl, Br pattern)detected; ¹H NMR (400 MHz, CD₃OD) δ 8.71 (s, 1H), 7.50 (d, 1H), 7.44 (s,1H), 7.20 (dd, 1H), 6.55 (d, 1H), 3.80 (s, 2H), 3.74 (d, 2H), 2.04 (s,6H), 1.18 (m, 1H), 0.51 (m, 2H), 0.27 (m, 2H).

4-[7-(4-Bromo-2-chlorophenylamino)-8-chloro-6-cyclopropylmethoxycarbamoylimidazo[1,2-a]pyridin-3-ylmethyl]-piperazine-1carboxylicacid tert-butyl ester

The reaction scheme for the synthesis of4-[7-(4-bromo-2-chlorophenylamino)-8-chloro-6-cyclopropylmethoxycarbamoylimidazo[1,2-a]pyridin-3-ylmethyl]-piperazine-1-carboxylicacid tert-butyl ester was similar to that to that described in Example18, using7-(4-bromo-2-chlorophenylamino)-8-chloroimidazo[1,2-a]pyridine-6-carboxylicacid (2-hydroxyethoxy)amide (33a) and piperazine-1-carboxylic acidtert-butyl ester to provide the desired product. MS APCI (+) m/z 669,671, 673 (M+, Cl, Br pattern) detected; ¹H NMR (400 MHz, CDCl₃) δ 8.80(s, 1H), 8.00 (s, 1H), 7.56 (d, 1H), 7.27 (dd, 1H), 6.67 (d, 1H), 4.54(s, 2H), 3.76 (d, 4H), 3.27 (m, 4H), 1.50 (s, 9H), 1.12 (m, 1H), 0.55(m, 2H), 0.28 (m, 2H).

7-(4-Bromo-2-chlorophenylamino)-8-chloro-3-(4-methylpiperazin-1-ylmethyl)-imidazo[1,2-a]pyridine-6-carboxylicacid cyclopropylmethoxyamide

The reaction scheme for the synthesis of7-(4-bromo-2-chlorophenylamino)-8-chloro-3-(4-methylpiperazin-1-ylmethyl)-imidazo[1,2-a]pyridine-6-carboxylicacid cyclopropylmethoxyamide was similar to that described in Example18, using7-(4-bromo-2-chlorophenylamino)-8-chloroimidazo[1,2-a]pyridine-6-carboxylicacid (2-hydroxyethoxy)amide (33a) and 1-methylpiperazine to provide thedesired product. MS APCI (+) m/z 581, 583, 585 (M+, Cl, Br pattern)detected; ¹H NMR (400 MHz, CDCl₃) δ 8.90 (s, 1H), 7.57 (s, 1H), 7.56 (d,1H), 7.22 (dd, 1H), 6.47 (d, 1H), 3.83 (s, 2H), 3.60 (d, 2H), 2.47 (m,8H), 2.31 (s, 3H), 1.02 (m, 1H), 0.56 (m, 2H), 0.26 (m, 2H).

7-(4-Bromo-2-fluorophenylamino)-8-fluoro-3-morpholin-4-ylmethylimidazo[1,2-a]pyridine-6-carboxylicacid ethoxyamide

The reaction scheme for the synthesis of7-(4-bromo-2-fluorophenylamino)-8-fluoro-3-morpholin-4-ylmethylimidazo[1,2-a]pyridine-6-carboxylicacid ethoxyamide is similar to that described in Example 18 using7-(4-bromo-2-fluorophenylamino)-8-fluoro-imidazo[1,2-a]pyridine-6-carboxylicacid ethoxyamide and morpholine to provide the desired product. MS ESI(+) m/z 510, 512 (M+, Br pattern) detected. ¹H NMR (400 MHz, CD₃OD) δ8.72 (s, 1H), 7.39 (s, 1H), 7.29 (dd, 1H), 7.17 (d, 1H), 6.76 (m, 1H),3.99 (q, 2H), 3.85 (s, 2H), 3.68 (m, 4H), 2.49 (br s, 4H), 1.29 (t, 3H).¹⁹F NMR (376 MHz, CD₃OD)−129.97 (s, 1F), −142.85 (s).

Example 19

Synthesis of7-(4-bromo-2-chlorophenylamino)-imidazo[1,2-a]pyridine-6-carboxylic acidcyclopropylmethoxyamide (44a)

Step A: Preparation of 4-(4-bromo-2-chlorophenylamino)-6-chloronicotinicacid tert-butyl ester (40): 2-tert-Butyl-1,3-diisopropylisourea (8.04 g,40.1 mmol) was added to a mixture of4-(4-bromo-2-chlorophenylamino)-6-chloronicotinic acid hydrochloridesalt (24) (2.91 g, 7.31 mmol) in THF (165 mL). After stirring for 2hours at room temperature and 30 minutes at reflux, the reaction mixturewas cooled to room temperature and diluted with EtOAc. The organic layerwas washed with 10% K₂CO₃ and brine, dried (Na₂SO₄) and concentrated.The resulting residue was dissolved in CH₂Cl₂ and filtered. The filtratewas concentrated and purified by flash column chromatography using theBiotage system (CH₂Cl₂) to give the desired product (40) (3.28 g, 78%).

Step B: Preparation of 6-azido-4-(4-bromo-2-chlorophenylamino)nicotinicacid tert-butyl ester (41): Sodium azide (1.51 g, 23.2 mmol) was addedto a suspension of 4-(4-bromo-2-chlorophenylamino)-6-chloronicotinicacid tert-butyl ester (40) (3.23 g, 7.73 mmol) in DMF (60 mL). Thereaction mixture was heated to 80° C. and stirred for 16 hours. Aftercooling to room temperature, the reaction was diluted with EtOAc andwashed with water, saturated NaHCO₃ and brine. The organic layer wasdried (Na₂SO₄) and concentrated. Purification by flash columnchromatography using the Biotage system (CH₂Cl₂) (repeated) provided thedesired product (41) (1.41 g, 43%).

Step C: Preparation of 6-amino-4-(4-bromo-2-chlorophenylamino)-nicotinicacid tert-butyl ester (42): Compound 42 was prepared as described inStep G of Example 9 using6-azido-4-(4-bromo-2-chlorophenylamino)nicotinic acid tert-butyl ester(41).

Step D: Preparation of7-(4-bromo-2-chlorophenylamino)imidazo[1,2-a]pyridine-6-carboxylic acid(43): Chloroacetaldehyde (12 μL, 0.188 mmol) was added to a mixture of6-amino-4-(4-bromo-2-chlorophenylamino)-nicotinic acid tert-butyl ester(42) (50 mg, 0.125 mmol) in EtOH (630 μL). After stirring the reactionmixture at 80° C. for 5 hours, an additional 12 μL of chloroacetaldehydewere added and heating was continued for 10 hours. The reaction mixturewas cooled to room temperature and diluted with EtOAc to give a cloudysemi-solution. The organic layer was washed with water, saturated NaHCO₃and brine. The organic layer contains a precipitate, which was collectedby filtration to give the desired product (43) (15 mg, 33%). MS APCI (−)m/z 364, 366, (M-, Cl, Br pattern) detected. ¹H NMR (400 MHz, DMSO-d₆) δ9.12 (s, 1H), 8.31 (s, 1H), 7.91 (s, 1H), 7.80 (s, 1H), 7.65-7.54 (m,2H), 6.91 (s, 1H).

Step E: Preparation of7-(4-bromo-2-chlorophenylamino)-imidazo[112-a]pyridine-6-carboxylic acidcyclopropylmethoxyamide (44a): Oxalyl chloride (2.0 M solution inCH₂Cl₂, 102 μL) was added to a stirred suspension of7-(4-bromo-2-chlorophenylamino)imidazo[1,2-a]pyridine-6-carboxylic acid(43) (15 mg, 0.041 mmol) in CH₂Cl₂ (1 mL) at 0° C. One drop of DMF wasadded. The reaction mixture was warmed to room temperature, stirred for25 minutes, and then concentrated. The residue was twice concentratedfrom toluene and dried in vacuo. The residue was suspended in CH₂Cl₂ (1mL), cooled to 0° C. and cyclopropylmethylhydroxylamine (36 mg, 0.409mmol) was added. The reaction mixture was warmed to room temperature,stirred for 2 hours and diluted with EtOAc. The organic layer was washedwith saturated NaHCO₃ and brine, dried (Na₂SO₄) and concentrated.Purification by flash column chromatography using the Biotage system(40:1 CH₂Cl₂/MeOH to 20:1 CH₂Cl₂/MeOH) provided the desired product(44a) as a tan solid (6 mg, 31%). MS APCI (−) m/z 433, 435 (M-, Cl, Brpattern) detected. ¹H NMR (400 MHz, CDCl₃) δ 8.68 (s, 1H), 7.69 (m, 1H),7.67 (d, 1H), 7.52-7.44 (m, 3H), 7.08 (s, 1H), 3.83 (d, 2H), 0.90 (m,1H), 0.62 (m, 2H), 0.35 (m, 2H).

The following compounds were synthesized in a similar manner asdescribed in Example 18, using the appropriate aniline in Step C.

7-(4-Bromo-2-fluorophenylamino)-imidazo[1,2-a]pyridine-6-carboxylic acid

MS ESI (+) m/z 350, 352 (M+, Br pattern) detected. (400 MHz, DMSO-d₆) δ9.13 (s, 1H), 7.94 (d, 1H). 7.70 (dd, 1H), 7.67 (d, 1H), 7.59 (t, 1H),7.47 (m, 1H), 6.84 (s, 1H). ¹⁹F NMR (376 MHz, DMSO-d₆) δ −128.9.

7-(4-Bromo-2-fluorophenylamino)-imidazo[1,2-a]pyridine-6-carboxylic acidcyclopropylmethoxyamide

MS ESI (+) m/z 419, 421 (M+, Br pattern) detected. ¹H NMR (400 MHz,DMSO-d₆) δ 11.91 (br s, 1H), 8.79 (s, 1H), 8.72 (br s, 1H), 7.81 (s,2H), 7.64 (m, 1H), 7.50 (m, 1H), 7.45 (m, 1H), 7.39 (m, 1H), 6.90 (s,1H), 3.74 (d, 2H), 1.14 (m, 1H), 0.55 (m, 2H), 0.29 (m, 2H). ¹⁹F NMR(376 MHz, DMSO-d₆) δ −124.3.

7-(4-bromo-2-fluorophenylamino)-imidazo[1,2-a]pyridine-6-carboxylic acid(2-hydroxyethoxy)-amide

MS ESI (+) m/z 409, 411 (M+, Br pattern) detected. ¹H NMR (400 MHz,DMSO-d₆) δ 12.02 (br s, 1H), 8.83 (s, 1H), 7.80 (s, 1H), 7.63 (s, 1H),7.51 (m, 1H), 7.45 (m, 1H), 7.39 (m, 1H), 6.91 (s, 1H), 4.79 (br s, 1H),3.94 (t, 2H), 3.64 (t, 2H). ¹⁹F NMR (376 MHz, DMSO-d₆) δ −124.4.

Example 20

Synthesis of7-(4-bromo-2-chlorophenylamino)-8-fluoroimidazo[1,2-a]pyridine-6-carboxylicacid cyclopropylmethoxyamide (47a)

Step A: Preparation of6-amino-4-(4-bromo-2-chlorophenylamino)-5-fluoronicotinic acidtert-butyl ester (45):1-Chloromethyl-4-fluoro-1,4-diazoniabicyclo[2.2.2]octanebis(tetrafluoroborate) (889 mg, 2.508 mmol) was added to a mixture of6-amino-4-(4-bromo-2-chlorophenylamino)nicotinic acid tert-butyl ester(42) (1.00 g, 2.51 mmol) in 1:1 MeOH/water (25 mL). After about 2 hours,the reaction mixture was diluted with EtOAc and water. The layers wereseparated and the organic layer washed with 0.5 N HCl and brine. Theaqueous washes were back extracted with EtOAc. The combined organicextracts were dried (Na₂SO₄) and concentrated. Purification by flashcolumn chromatography using the Biotage system (20:1 hexanes/EtOAc to15:1 hexanes/EtOAc) provided the desired product (45) as a yellow solid(75 mg, 7%).

Step B: Preparation of7-(4-bromo-2-chlorophenylamino)-8-fluoroimidazo[1,2-a]pyridine-6-carboxylicacid (46): Chloroacetaldehyde (23 μL, 0.360 mmol) was added to a mixtureof 6-amino-4-(4-bromo-2-chlorophenylamino)-5-fluoronicotinic acidtert-butyl ester (45) (75 mg, 0.180 mmol) in EtOH (1 mL). After stirringthe reaction mixture at 70° C. for 10 hours, an additional 10 μLchloroacetaldehyde was added and heating was continued for 33 hours. Thereaction mixture was cooled to room temperature and desired product (46)was collected by filtration. The filtrate was diluted with EtOAc andwashed with water, saturated NaHCO₃ and brine. The organic layer wasdried (Na₂SO₄) and concentrated to give additional product (46) (51 mg,74% combined recovery). MS APCI (−) m/z 382, 384, 386 (M-, Cl, Brpattern) detected. ¹H NMR (400 MHz, CD₃OD) δ 8.86 (s, 1H), 7.85 (s, 1H),7.53 (s, 2H), 7.32 (d, 1H), 6.82 (t, 1H). ¹⁹F NMR (376 MHz, CD₃OD)−148.3(s).

7-(4-Bromo-2-chlorophenylamino)-8-fluoroimidazo[1,2-a]pyridine-6-carboxylicacid (46) was alternatively synthesized by the route shown below.

In the first step of this alternative procedure,4,6-dichloro-5-fluoronicotinic acid (J. Heterocyclic Chemistry 1993, 30,855-859) was used to synthesize4-(4-bromo-2-chlorophenylamino)-5-bromo-6-chloronicotinic acid accordingto the alternate procedure described in Example 9, Step B.

Step C: Preparation of7-(4-bromo-2-chlorophenylamino)-8-fluoroimidazo[1,2-a]pyridine-6-carboxylicacid cyclopropylmethoxyamide (47a): Compound 47 was prepared asdescribed in Step E of Example 19 using7-(4-bromo-2-chlorophenylamino)-8-fluoroimidazo[1,2-a]pyridine-6-carboxylicacid (46) to give 15 mg (24%) of the desired product (47a) as a whitesolid. MS APCI (+) m/z 453, 455, 457 (M+, Cl, Br pattern) detected. ¹HNMR (400 MHz, CD₃OD) δ 8.66 (s, 1H), 7.93 (m, 1H), 7.61 (s, 1H), 7.56(d, 1H), 7.32 (dd, 1H), 6.73 (q, 1H) 3.70 (d, 2H), 1.14 (m, 1H), 0.56(m, 2H), 0.26 (m, 2H): ¹⁹F (400 MHz, CD₃OD)−139.4 (s).

The following compounds were synthesized in a similar manner asdescribed in Example 20.

7-(4-Bromo-2-chlorophenylamino)-8-fluoroimidazo[1,2-a]pyridine-6-carboxylicacid (2-hydroxyethoxy)amide

MS APCI (+) m/z 443, 445, 447 (M+, Cl, Br pattern) detected; ¹H NMR (400MHz, CD₃OD) δ 8.69 (s, 1H), 7.89 (m, 1H), 7.59 (s, 1H), 7.55 (d, 1H),7.31 (dd, 1H), 6.72 (q, 1H), 4.01 (t, 2H), 3.76 (t, 2H); ¹⁹F (400 MHz,CD₃OD)−139.7 (s).

7-(4-Bromo-2-fluorophenylamino)-8-fluoroimidazo[1,2-a]pyridine-6-carboxylicacid

MS ESI (+) m/z 368, 370 (M+, Br pattern) detected. ¹H NMR (400 MHz,DMSO-d₆) δ 9.21 (s, 1H), 8.08 (m, 1H), 7.66 (s, 1H), 7.55 (dd, 1H), 7.29(d, 1H), 6.92 (m, 1H). ¹⁹F (376 MHz, DMSO-d₆)−127.9 (s, 1F), −141.1 (s,1F).

7-(4-Bromo-2-fluorophenylamino)-8-fluoroimidazo[1,2-a]pyridine-6-carboxylicacid cyclopropylmethoxyamide

MS ESI (+) m/z 437, 439 (M+, Br pattern) detected. ¹H NMR (400 MHz,CD₃OD) δ 9.55 (br s, 1H), 8.57 (s, 1H). 7.68 (s, 2H), 7.65 (s, 1H), 7.28(m, 1H), 7.14 (m, 1H), 6.78 (br s, 1H), 6.63 (m, 1H), 3.72 (m, 2H), 1.07(m, 1H), 0.59 (m, 2H), 0.28 (m, 2H). ¹⁹F NMR (376 MHz, CD₃OD) δ−128.9(s, 1F), −138.1 (s, 1F).

7-(4-Bromo-2-methylphenylamino)-8-fluoroimidazo[1,2-a]pyridine-6-carboxylicacid

MS APCI (+) m/z 364, 366 (M+, Br pattern) detected. ¹H NMR (400 MHz,DMSO-d₆) δ 9.22 (s, 1H), 8.07 (m, 1H), 7.68 (s, 1H), 7.42 (d, 1H), 7.28(dd, 1H), 6.78 (t, 1H), 2.29 (s, 3H). ¹⁹F (376 MHz, DMSO-d₆)−142.5 (s).

7-(2,4-dichlorophenylamino)-8-fluoroimidazo[1,2-a]pyridine-6-carboxylicacid

MS APCI (+) m/z 440, 442 (M+, Cl pattern) detected. ¹H NMR (400 MHz,DMSO-d₆) δ 9.27 (s, 1H), 8.13 (s, 1H), 7.71 (s, 1H), 7.62 (s, 1H), 7.32(dd, 1H), 6.95 (t, 1H). ¹⁹F (376 MHz, DMSO-d₆)−129.9 (s).

8-Fluoro-7-(2-fluoro-4-methylphenylamino)-imidazo[1,2-a]pyridine-6-carboxylicacid

MS APCI (+) m/z 304 (M+1) detected. ¹H NMR (400 MHz, DMSO-d₆) δ 9.48 (brs, 1H), 9.32 (s, 1H), 8.12 (s, 1H), 7.86 (s, 1H), 7.12 (m, 2H), 6.98 (d,1H), 2.31 (s, 3H). ¹⁹F (376 MHz, DMSO-d₆)−128.1 (s, 1F), −148.8 (s, 1F).

7-(4-Chloro-2-fluorophenylamino)-8-fluoroimidazo[1,2-a]pyridine-6-carboxylicacid

MS APCI (+) m/z 324, 326 (M+, Cl pattern) detected. ¹H NMR (400 MHz,DMSO-d₆) δ 9.19 (s, 1H), 8.07 (m, 1H), 7.64 (s, 1H), 7.45 (dd, 1H), 7.17(d, 1H), 6.98 (m, 1H). ¹⁹F (376 MHz, CD₃OD)−128.8 (s, 1F), −154.8 (s,1F).

Example 213-Chloro-7-(2,4-dichlorophenylamino)-8-fluoroimidazo[1,2-a]pyridine-6-carboxylicacid (2-hydroxyethoxy)-amide

Step A: Preparation of3-chloro-7-(2,4-dichlorophenylamino)-8-fluoroimidazo[1,2a]pyridine-6-carboxylicacid methyl ester.7-(2,4-Dichlorophenylamino)-8-fluoroimidazo[1,2-a]pyridine-6-carboxylicacid methyl ester (57.0 mg, 0.16 mmol), which was synthesized in amanner similar to the alternate procedure described in Example 20, StepB, was dissolved into DMF (3 mL). Added N-chlorosuccinimide (17.0 mg,0.13 mmol) and HCl (1.0 M aqueous solution, 16 μL, 0.016 mmol). Afterstirring for 16 hours, the suspension was diluted with ethyl acetate,washed with NaHSO₃, water (2×), brine, dried over Na₂SO₄ andconcentrated to a yellow solid. Purification by flash columnchromatography (4:1 hexanes/ethyl acetate) provided the desired productas a light yellow solid (40 mg, 64%). MS APCI (+) m/z 388, 390 (M+, Clpattern) detected. ¹H NMR (400 MHz, CDCl₃) δ 8.70 (s, 2H), 7.58 (s, 1H),7.40 (d, 1H), 7.14 (dd, 1H), 6.77 (t, 1H), 4.02 (s, 3H): ¹⁹F NMR (376MHz, CDCl₃)−135.84 (s).

Step B: Preparation of3-chloro-7-(2,4-dichlorophenylamino)-8-fluoroimidazo[1,2-a]pyridine-6-carboxylicacid (2-vinyloxyethoxy)-amide. LiHMDS (1.0 M in hexanes, 0.34 mL, 0.34mmol) was added to a solution3-chloro-7-(2,4-dichlorophenylamino)-8-fluoroimidazo[1,2-a]pyridine-6-carboxylicacid methyl ester (38 mg, 0.098 mmol) andO-(2-vinylox-ethyl)-hydroxylamine (25 mg, 0.24 mmol) in THF (1.0 mL)cooled to 0° C. The solution was allowed to warm to room temperature andstir for 16 hours. The solution was diluted with ethyl acetate, washedwith saturated NaHCO₃, water (3×), brine, dried over Na₂SO₄ andconcentrated to a yellow liquid. Purification by flash columnchromatography (20:1 to 10:1 dichloromethane/MeOH) provided the desiredproduct as a yellow solid (42 mg, 93%). MS APCI (+) m/z 459, 461 (M+, Clpattern) detected.

Step C: Preparation of3-chloro-7-(2,4-dichlorophenylamino)-8-fluoroimidazo[1,2-a]pyridine-6-carboxylicacid (2-hydroxyethoxy)-amide.3-Chloro-7-(2,4-dichlorophenylamino)-8-fluoro-imidazo[1,2-a]pyridine-6-carboxylicacid (2-vinyloxy-ethoxy)-amide was converted to the desired productaccording to Example 3, Step B, to provide 31 mg (78%) of desiredproduct as a light yellow solid. MS APCI (+) m/z 433, 435 (M+, Clpattern) detected. ¹H NMR (400 MHz, CD₃OD) δ 8.52 (s, 1H), 7.57 (s, 1H),7.42 (d, 1H), 7.16 (dd, 1H), 6.79 (dd, 1H), 4.07 (m, 2H), 3.80 (m, 2H).¹⁹F NMR (376 MHz, CD₃OD) −140.83 (s).

Example 227-(4-Ethyl-2-fluorophenylamino)-8-fluoroimidazo[1,2-a]pyridine-6-carboxylicacid ethoxyamide

Step A: Preparation of8-fluoro-7-(2-fluoro-4-vinylphenylamino)-imidazo[1,2-a]pyridine-6-carboxylicacid methyl ester.8-Fluoro-7-(2-fluoro-4-iodophenylamino)-imidazo[1,2-a]pyridine-6-carboxylicacid methyl ester (250 mg, 0.58 mmol), which was synthesized in asimilar manner to the alternative synthesis set forth in Example 20 StepB, was suspended in isopropyl alcohol (6 mL) and tetrahydrofuran (1 mL).Potassium vinyltrifluoroborate (90 mg, 0.67 mmol) and triethylamine(0.165 mL, 1.2 mmol) were added, at which time the reaction mixturebecame a solution. The reaction mixture was sparged with N₂.PdCl₂(dppf)-CH₂Cl₂ (2 mol %, 9 mg) was then added and the reaction washeated to 90° C. and stirred under N₂ for 16 hours. The reaction mixturewas cooled to room temperature and diluted with water, followed byextraction with ethyl acetate (2×). The combined organic layers werewashed with saturated aqueous NaCl, dried over Na₂SO₄ and concentrated.Purification of the crude product by flash column chromatography (30:1dichloromethane/methanol) provided the desired product (143 mg, 81%) asa dark yellow solid. MS ESI (+) m/z 330 (M+1) detected.

Step B: Preparation of7-(4-ethyl-2-fluorophenylamino)-8-fluoroimidazo[1,2-a]pyridine-6-carboxylicacid methyl ester.8-Fluoro-7-(2-fluoro-4-vinyl-phenylamino)-imidazo[1,2-a]pyridine-6-carboxylic acid methyl ester (165 mg, 0.50 mmol) was suspended inethanol (5mL), added 10% Pd/C (267 mg, 0.25 mmol) and placed under anatmosphere of H₂. The reaction mixture was stirred vigorously at roomtemperature for 16 hours. The reaction mixture was filtered throughCELITE, washed with ethanol and tetrahydrofuran and the filtrateconcentrated to a yellow oil. Purification of the crude product by flashcolumn chromatography (gradient of dichloromethane to 20:1dichloromethane/methanol) provided the desired product (110 mg, 66%) asa dark yellow solid. MS ESI (+) m/z 332 (M+1) detected.

Step C: Preparation of7-(4-ethyl-2-fluorophenylamino)-8-fluoroimidazo[1,2-a]pyridine-6-carboxylicacid ethoxyamide.7-(4-Ethyl-2-fluorophenylamino)-8-fluoroimidazo[1,2-a]pyridine-6-carboxylicacid methyl ester (40 mg, 0.12 mmol) was converted to the desiredproduct according to procedure of Step B of Example 21, usingO-ethyl-hydroxylamine HCl salt, to provide the desired product as ayellow colored solid (31 mg, 71%). MS ESI (+) m/z 361 (M+1) detected. ¹HNMR (400 MHz, CD₃OD) δ 8.62 (s, 1H), 7.86 (s, 1H), 7.56 (s, 1H), 6.97(d, 1H), 6.90-6.80 (m, 2H), 3.88 (q, 2H), 2.60 (q, 2H), 1.23 (m, 9H).¹⁹F (376 MHz, CD₃OD)−132.7 (s, 1F), −145.2 (s, 1F).

Example 233-Ethyl-8-fluoro-7-(2-fluoro-4-methylphenylamino)-imidazo[1,2-a]pyridine-6-carboxylicacid (2-hydroxyethoxy)-amide

Step A: Preparation of8-Fluoro-7-(2-fluoro-4-methylphenylamino)-3-iodo-imidazo[1,2-a]pyridine-6-carboxylicacid methyl ester. N-iodosuccinimide (134 mg, 0.60 mmol) was added in asingle portion to a solution of8-fluoro-7-(2-fluoro-4-methylphenylamino)-imidazo[1,2-a]pyridine-6-carboxylicacid methyl ester (172 mg, 0.54 mmol) in acetonitrile (10 mL) resultingin a thick precipitate after ten minutes of stirring. The suspension wasdiluted with ethyl acetate, washed with NaHSO₃, saturated NaHCO₃, water,brine, and dried over Na₂SO₄. Concentration provided the desired productas a yellow solid (241 mg, 100%). MS APCI (+) m/z 444 (M+1) detected. ¹HNMR (400 MHz, CD₃OD) δ 8.78 (s, 1H), 7.62 (s, 1H), 6.95 (m, 3H), 4.00(s, 3H), 2.33 (s, 3H). ¹⁹F NMR (376 MHz, CD₃OD)−131.11 (s, 1F), −146.71(s, 1F).

Step B: Preparation of8-fluoro-7-(2-fluoro-4-methylphenylamino)-3-trimethylsilanylethynyl-imidazo[1,2-a]pyridine-6-carboxylicacid methyl ester. THF (1.0 mL), diisopropylamine (95 μL, 0.68 mmol) andtrimethylsilylacetylene (38 μL, 0.27 mmol) were added to a mixture of8-fluoro-7-(2-fluoro-4-methylphenylamino)-3-iodo-imidazo[1,2-a]pyridine-6-carboxylicacid methyl ester (100 mg, 0.23 mmol), CuI (4.0 mg, 0.023 mmol) andPdCl₂(PPh₃)₂ (16 mg, 0.023 mmol). The solution was stirred at roomtemperature for 16 hours. The reaction was diluted with ethyl acetate,washed with saturated NaHCO₃ (3×), brine (2×), dried over Na₂SO₄ andconcentrated to a dark brown solid. Flash column chromatography (40:1dichloromethane/MeOH) provide desired product (30 mg, 32%) as a yellowliquid. MS APCI (+) m/z 414 (M+1) detected. ¹H NMR (400 MHz, CDCl₃) δ8.77 (s, 1H), 8.61 (s, 1H), 7.80 (s, 1H), 6.90 (m, 3H), 4.00 (s, 3H),2.32 (s, 3H), 0.33 (s, 9H). ¹⁹F NMR (376 MHz, CDCl₃)−129.12 (s, 1F),−141.99 (s, 1F).

Step C: Preparation of3-ethynyl-8-fluoro-7-(2-fluoro-4-methylphenylamino)-imidazo[1,2-a]pyridine-6-carboxylicacid methyl ester. Potassium carbonate (70 mg, 0.51 mmol) was added to asolution of8-fluoro-7-(2-fluoro-4-methylphenylamino)-3-trimethylsilanylethynylimidazo[1,2-a]pyridine-6-carboxylicacid methyl ester in MeOH (5 mL) and stirred at room temperature for twohours. The mixture was diluted with ethyl acetate, washed with brine(2×), dried over Na₂SO₄ and concentrated to a brown residue. Flashcolumn chromatography (dichloromethane to 80:1 dichloromethane/MeOH)provided the desired product (16 mg, 65%) as a yellow liquid. MS APCI(+) m/z 342 (M+1) detected.

Step D: Preparation of3-ethyl-8-fluoro-7-(2-fluoro-4-methylphenylamino)-imidazo[1,2-a]pyridine-6-carboxylicacid methyl ester. A mixture of3-ethynyl-8-fluoro-7-(2-fluoro-4-methylphenylamino)-imidazo[1,2-a]pyridine-6-carboxylicacid methyl ester (16 mg, 0.047 mmol) and 10% Pd/C (5 mg) under anatmosphere of hydrogen (balloon) was stirred vigorously for one hour.The mixture was filtered through a plug of rinsing with dichloromethaneand concentrated to provide the desired product (14 mg, 86%) as a yellowfoam. MS APCI (+) m/z 346 (M+1) detected. ¹H NMR (400 MHz, CDCl₃) δ 8.51(s, 1H), 8.31 (s, 1H), 7.38 (s, 1H), 6.91 (d, 1H), 6.84 (s, 2H), 3.97(s, 3H), 2.86 (q, 2H), 2.30 (s, 3H), 1.43 (t, 3H).

Step E: Preparation of3-ethyl-8-fluoro-7-(2-fluoro-4-methylphenylamino)-imidazo[1,2-a]pyridine-6-carboxylicacid (2-hydroxy-ethoxy)-amide.3-Ethyl-8-fluoro-7-(2-fluoro-4-methyl-phenylamino)-imidazo[1,2-a]pyridine-6-carboxylicacid methyl ester (14 mg, 0.041 mmol) was converted to the desiredproduct according to the Step A of example 21 and Step B of Example 3,Step B to provide the desired product (hydrochloride salt) as a tancolored solid (9 mg, 51% over two steps). MS APCI (+) m/z 391 (M+1)detected. ¹H NMR (400 MHz, CD₃OD) δ 8.63 (s, 1H), 7.65 (s, 1H), 7.16 (m,1H), 7.01 (m, 2H), 4.04 (br s, 2H), 3.79 (br s, 2H), 2.97 (q, 2H), 2.35(2, 3H), 1.45 (t, 3H). ¹⁹F NMR (376 MHz, CD₃OD)−127.69 (s, 1F), −155.07(s, I F).

Example 24

Synthesis of7-(4-bromo-2-chlorophenylamino)-8-chloro-3-methyl-[1,2,4]-triazolo-[4,3-a]pyridine-6-carboxylicacid cyclopropylmethoxyamide (53a)

Step A: Preparation of4-(4-bromo-2-chlorophenylamino)-5-chloro-6-hydrazinonicotinic acid ethylester (49): 4-(4-Bromo-2-chlorophenylamino)-5,6-dichloronicotinic acidethyl ester (48) was prepared by standard methods from4-(4-bromo-2-chlorophenylamino)-5,6-dichloronicotinic acid. Hydrazinemonohydrate (0.59 mL, 12.16 mmol) was added to a solution of4-(4-bromo-2-chlorophenylamino)-5,6-dichloronicotinic acid ethyl ester(48) (1.72 g, 4.05 mmol) in N,N-dimethylacetamide (20 mL). Afterstirring at 90° C. for 1 hour, the reaction mixture was cooled to roomtemperature and diluted with EtOAc. The organic layer was washed withwater and brine, dried (Na₂SO₄) and concentrated. Purification by flashcolumn chromatography using the Biotage system (20:1 CH₂Cl₂:EtOAc)provided the desired product (49) (307 mg, 18%).

Step B: Preparation of7-(4-bromo-2-chlorophenylamino)-8-chloro-3-methyl-[1,2,4]triazolo[4,3-a]pyridine-6-carboxylicacid ethyl ester (51a): Acetic anhydride (22 μL, 0.238 mmol) was addedto a solution of4-(4-bromo-2-chlorophenylamino)-5-chloro-6-hydrazinonicotinic acid ethylester (49) (0.100 g, 0.238 mmol) and triethylamine (66 μL, 0.476 mmol)in CH₂Cl₂ (2.5 mL) at 0° C., and then the solution was warmed to roomtemperature to provide compound 50a (not isolated). After 10 minutes,POCl₃ (87 μL, 0.952 mmol) was added dropwise and the reaction mixturewas warmed to room temperature. After 16 hours, the reaction mixture washeated to reflux and stirred for 3 days. The reaction mixture was cooledto room temperature and concentrated. The residue was diluted with EtOAcand saturated NaHCO₃ was added and the mixture stirred for 20 minutes.The layers were separated and the organic layer was washed with brine.The aqueous washings were back extracted with EtOAc. The combinedorganic extracts were dried (Na₂SO₄) and concentrated. Purification byflash column chromatography using the Biotage system (9:1 CH₂Cl₂/EtOAc)provided compound 51a (80 mg, 75%).

Step C: Preparation of7-(4-bromo-2-chlorophenylamino)-8-chloro-3-methyl-[1,2,4]triazolo[4,3-a]pyridine-6-carboxylicacid (52a): Sodium hydroxide (715 μL of a 1 M solution) was added to asolution of7-(4-bromo-2-chlorophenylamino)-8-chloro-3-methyl-[1,2,4]triazolo[4,3-a]pyridine-6-carboxylicacid ethyl ester (51a) (79 mg, 179 mmol) in 3:1 THF:water (4.5 mL).After 16 hours, the reaction mixture was poured into a separatoryfunnel, diluted with brine and acidified with 1 M HCl to about pH 2. Theaqueous layer was extracted with 1:1 EtOAc/THF. The combined organicextracts were dried (Na₂SO₄) and concentrated and the residue (52a) wascarried forward without further purification.

Step D: Preparation of7-(4-bromo-2-chlorophenylamino)-8-chloro-3-methyl-[1,2,4]triazolo[4,3-a]pyridine-6-carboxylicacid cyclopropylmethoxyamide (53a): Compound 53a was prepared asdescribed in Example 2 using7-(4-bromo-2-chlorophenylamino)-8-chloro-3-methyl-[1,2,4]triazolo[4,3-a]pyridine-6-carboxylicacid (52a) to give 2 mg (5%) of the desired product. MS APCI (−) m/z482, 484, 486 (M-, Cl, Br pattern) detected.

Example 25

Synthesis of7-(4-bromo-2-chlorophenylamino)-8-chloro-3-methyl-[1,2,4]triazolo[4,3-a]pyridine-6-carboxylicacid (2-hydroxyethoxy)-amide (54a)

Compound 54a was prepared as described herein starting with7-(4-bromo-2-chlorophenylamino)-8-chloro-3-methyl-[1,2,4]triazolo[4,3-a]pyridine-6-carboxylicacid (52a) to give 1 mg (2% for the two steps) desired product (54a). MSAPCI (−) m/z 472, 474, 476 (M-, Cl, Br pattern) detected.

Example 26

Synthesis of3-benzyl-7-(4-bromo-2-chlorophenylamino)-8-chloro-[1,2,4]triazolo[4,3-a]pyridine-6-carboxylicacid cyclopropylmethoxyamide (53b)

Step A: Preparation of3-benzyl-7-(4-bromo-2-chlorophenylamino)-8-chloro-[1,2,4]triazolo[4,3-a]pyridine-6-carboxylicacid methyl ester (51b): Phenylacetyl chloride (152 μL, 1.148 mmol) wasadded to a solution of4-(4-bromo-2-chlorophenylamino)-5-chloro-6-hydrazinonicotinic acidmethyl ester (49) (0.233 g, 0.574 mmol) and triethylamine (160 μL, 1.148mmol) in CH₂Cl₂ (5.7 mL) at 0° C. After warming to room temperature, anadditional 75 μL phenylacetyl chloride was added. After 6 hours, thereaction mixture was concentrated and diluted with EtOAc. The organiclayer was washed with water and brine, dried (Na₂SO₄) and concentrated.The residue (50b) was diluted with dichloroethylene (2 mL) and POCl₃(465 μL, 5.082 mmol) was added. After stirring at reflux for 12 hours,the reaction mixture was cooled to room temperature and concentrated.The residue was diluted with EtOAc and saturated NaHCO₃ was added andthe mixture stirred for 20 minutes. The resulting solid was collected byfiltrate to give the desired product (51b) (97 mg, 30%).

Step B: Preparation of3-benzyl-7-(4-bromo-2-chlorophenylamino)-8-chloro-[1,2,4]triazolo[4,3-a]pyridine-6-carboxylicacid cyclopropylmethoxyamide (53b): Compound 53b was prepared asdescribed in Step C of Example 24 and Example 2 using3-benzyl-7-(4-bromo-2-chlorophenylamino)-8-chloro-[1,2,4]triazolo[4,3-a]pyridine-6-carboxylicacid methyl ester (5 lb) as the starting material to give 5 mg (4% forthe two steps) of the desired product (53b). MS APCI (−) m/z 558, 560,562 (M-, Cl, Br pattern) detected.

¹H NMR (400 MHz, CDCl₃) δ 8.22 (s, 1H), 7.30 (m, 6H), 6.50 (d, 1H), 4.53(s, 2H), 3.49 (m, 2H), 0.94 (m, 1H), 0.51 (m, 2H), 0.19 (m, 2H).

Example 27

Synthesis of 6-(2-chlorophenylamino)-7-fluoro-3-methylbenzo[c]isoxazole-5-carboxylic acid (56)

Step A: Preparation of6-(2-chlorophenylamino)-7-fluoro-3-methyl-benzo[c]-isoxazole-5-carboxylicacid methyl ester (55): Sodium azide (128 mg, 1.95 mmol) was added to amixture of 5-acetyl-2-(2-chlorophenylamino)-3,4-difluorobenzoic acidmethyl ester (6) (601 mg, 1.59 mmol) in 3:1 acetone:water (16 ml) andheated to reflux. After 16 hours, the reaction mixture was cooled toroom temperature, and diluted with EtOAc and water. The organic layerwas washed with water, dried (MgSO₄) and concentrated. The resultingresidue was diluted with water (8 mL) and heated to reflux. After 5hours, the mixture was cooled to room temperature and diluted withEtOAc. The organic layer was washed with water, dried (MgSO₄) andconcentrated. Purification by flash column chromatography using theBiotage system (20% EtOAc in hexanes) provided the desired product (55)(410 mg, 77%).

Step B: Preparation of 6-(2-chlorophenylamino)-7-fluoro-3-methylbenzo[c]-isoxazole-5-carboxylic acid (56): To a solution of6-(2-chlorophenylamino)-7-fluoro-3-methylbenzo[c]-isoxazole-5-carboxylicacid methyl ester (55) (100 mg, 0.299 mmol) in 6:1 THF:water (3.5 mL)was added LiOH (0.60 ml of a 1 M solution in water). After 1 hour, thereaction was acidified to pH 1 with 1 N HCl, diluted with water andextracted with EtOAc. The combined organic extracts were washed withwater, dried (MgSO₄) and concentrated to give the desired product (56)(87 mg, 91%). MS APCI (−) m/z 319, 321 (M+, Cl pattern) detected. ¹H NMR(400 MHz, CD₃OD) δ 8.45 (s, 1H), 7.38 (dd, 1H), 7.20 (m, 1H), 6.91 (m,2H), 2.88 (s, 3H): ¹⁹F NMR (376 MHz, CD₃OD)−136.40 (s).

Example 28

Synthesis of 6-(2-chlorophenylamino)-7-fluoro-3-methylbenzo[c]isoxazole-5-carboxylic acid (2-hydroxyethoxy)amide (57a)

Compound 57a was prepared as described in Example 27 using6-(2-chlorophenylamino)-7-fluoro-3-methylbenzo[c]isoxazole-5-carboxylicacid (56) to give 35 mg (44%) desired product. MS APCI (−) m/z 388, 390(M+, Cl pattern) detected; ¹H NMR

MHz, CD₃OD) δ 7.73 (s, 1H), 7.36 (d, 1H), 7.17 (t, 1H), 6.89 (t, 1H),6.81 (dd, 1H), 3.72(d, 2H), 2.87 (s, 3H), 1.15 (m, 1H), 0.54 (d, 2H),0.26 (d, 2H); ¹⁹F NMR (376 MHz, CD₃OD)−135.08 (s).

Example 29

Synthesis of6-(2-chloro-phenylamino)-7-fluoro-3-methylbenzo[c]isoxazole-5-carboxylicacid cyclopropylmethoxy-amide (57b)

Compound 57b was prepared as described in Example 27 using6-(2-chlorophenylamino)-7-fluoro-3-methylbenzo[c]isoxazole-5-carboxylicacid (56) to give 35 mg (44%) desired product. MS APCI (−) m/z 388, 390(M+, Cl pattern) detected; ¹H NMR

MHz, CD₃OD) δ 7.73 (s, 1H), 7.36 (d, 1H), 7.17 (t, 1H), 6.89 (t, 1H),6.81 (dd, 1H), 3.72(d, 2H), 2.87 (s, 3H), 1.15 (m, 1H), 0.54 (d, 2H),0.26 (d, 2H); ¹⁹F NMR (376 MHz, CD₃OD)−135.08 (s).

Example 30

Synthesis of7-(4-bromo-2-chlorophenylamino)-8-chloro-3-methylaminomethyl-imidazo[1,2-a]pyridine-6-carboxylicacid cyclopropylmethoxyamide (63)

Step A: Preparation of7-(4-bromo-2-chlorophenylamino)-8-chloro-3-formylimidazo[1,2-a]pyridine-6-carboxylicacid methyl ester (58): A suspension of6-amino-4-(4-bromo-2-chlorophenylamino)-5-chloro-nicotinic acid methylester (28) (1.06 g, 2.72 mmol) and 2-chloro-malonaldehyde (587 mg, 5.43mmol) was heated to 80° C. for 45 minutes. The solution was allowed tocool to room temperature, and then washed with saturated aqueous NaHCO₃,and brine. The organic layer was dried over NaSO₄, filtered,concentrated in vacuo, and purified by column chromatography (20:1methylene chloride/methanol) to give the desired product as a darkyellow solid. The solid was triturated with ethyl acetate and isolatedby filtration to provide the desired product as a yellow solid (0.436 g,36%). MS (APCI+) m/z 442, 444, 446 (M+; Cl, Br pattern) detected.

Step B: Preparation of7-(4-bromo-2-chlorophenylamino)-8-chloro-3-methylaminomethylimidazo[1,2-a]pyridine-6-carboxylicacid methyl ester (59): A suspension of7-(4-bromo-2-chlorophenylamino)-8-chloro-3-formylimidazo[1,2-a]pyridine-6-carboxylicacid methyl ester (58) (25 mg, 0.056 mmol), acetic acid (7 μL, 0.11mmol), and methylamine (2.0 M solution in THF, 56 μL, 0.11 mmol) wasstirred for 0.5 hours. Sodium triacetoxyborohydride (36 mg, 0.17 mmol)was added and the solution allowed to stir overnight. The reactionmixture was concentrated to dryness and purified by flash columnchromatography (dichloromethane followed by 10:1dichloromethane/methanol) to provide the desired product as a yellowsolid (12 mg, 46%). MS (APCI+) m/z 455, 457, 459 (M+; Cl, Br pattern)detected.

Step C: Preparation of7-(4-bromo-2-chlorophenylamino)-3-[(tert-butoxycarbonylmethylamino)-methyl]-8-chloroimidazo[1,2-a]pyridine-6-carboxylicacid methyl ester (60): Di-tert-butyl dicarbonate (6 mg, 0.029 mmol) andtriethylamine (4 μL, 0.029 mmol) were added to a solution of7-(4-bromo-2-chlorophenylamino)-8-chloro-3-methyaminomethylimidazo[1,2-a]pyridine-6-carboxylicacid methyl ester (59) (12 mg, 0.026 mmol) in dichloromethane. Thesolution was stirred at room temperature for 0.5 hr after which timeHPLC analysis indicated the reaction had gone to completion. Thesolution was rotovapped to dryness to provide the desired product as ayellow foam (15 mg, quantitative). MS (APCI+) m/z 557, 559, 561 (M+; Cl,Br pattern) detected.

Step D: Preparation of7-(4-bromo-2-chlorophenylamino)-3-[(tert-butoxycarbonylmethylamino)-methyl]-8-chloroimidazo[1,2-a]pyridine-6-carboxylicacid (61): Sodium hydroxide (1.0 M aqueous solution, 0.16 mL, 0.16 mmol)was added to a solution of7-(4-bromo-2-chlorophenylamino)-3-[(tert-butoxycarbonylmethylamino)-methyl]-8-chloroimidazo[1,2-a]pyridine-6-carboxylicacid methyl ester (60) (15 mg, 0.026 mmol) in 4:1 MeOH/water (5 mL).When the reaction was complete, the solution was diluted with water,acidified to pH 3 by addition of 1.0 M aqueous HCl, and extracted withethyl acetate. The organic extracts were dried over NaSO₄, filtered,concentrated in vacuo to provide the desired product as a whitecrystalline solid (12 mg, 84%). MS (APCI−) m/z 541, 543, 545 (M-; Cl, Brpattern) detected.

Step E: Preparation of[7-(4-bromo-2-chlorophenylamino)-8-chloro-6-cyclopropylmethoxycarbamoylimidazo[1,2-a]pyridin-3-ylmethyl]-methylcarbamicacid tert-butyl ester (62): EDCI (6 mg, 0.033 mmol) and HOBt (5 mg,0.033 mmol) were added to a solution of7-(4-bromo-2-chlorophenylamino)-3-[(tert-butoxycarbonylmethylamino)-methyl]-8-chloroimidazo[1,2-a]pyridine-6-carboxylicacid (61) in dimethylacetamide (0.4 mL). The yellow solution was allowedto stir at room temperature for 0.5 hours after which timeO-cyclopropylmethyl-hydroxylamine (6 mg, 0.066 mmol) and triethylamine(6 μL, 0.044 mmol) were added and the solution allowed to stirovernight. The reaction mixture was diluted with ethyl acetate, washedwith water, saturated aqueous ammonium chloride, saturated aqueouspotassium carbonate and brine. The organic phase was dried over NaSO₄,filtered, concentrated in vacuo to provide the desired product as ayellow residue (11.5 mg, 85%). MS (APCI+) m/z 612, 614, 616 (M+; Cl, Brpattern) detected.

Step F: Preparation of7-(4-bromo-2-chlorophenylamino)-8-chloro-3-methylaminomethylimidazo[1,2-a]pyridine-6-carboxylicacid cyclopropylmethoxyamide (63): A solution of[7-(4-bromo-2-chlorophenylamino)-8-chloro-6-cyclopropylmethoxy-carbamoyl-imidazo[1,2-a]pyridin-3-ylmethyl]-methyl-carbamicacid tert-butyl ester (62) in 1:1 dichloromethane/trifluoroacetic acidwas stirred for two hours. Solvent was removed under reduced pressureand the residue redissolved into ethyl acetate. The organic solution waswashed with saturated aqueous potassium carbonate and brine. The aqueouswashes were back-extracted with ethyl acetate. The combined organicextracts are dried over NaSO₄, filtered, and concentrated in vacuo toprovide the desired product as a yellow solid (8 mg, 83%). MS (APCI+)m/z 512, 514, 516 (M+; Cl, Br pattern) detected. ¹H NMR (400 MHz,methanol-d₄) δ 8.72 (s, 1H), 7.58 (s, 1H), 7.54 (s, 1H), 7.25 (d, 1H),6.55 (d, 1H), 4.23 (s, 2H), 3.67 (d, 2H), 2.51 (s, 3H), 1.13 (m, 1H),0.50 (d, 2H), 0.24 (d, 2H).

Example 31

6-(4-bromo-2-chlorophenylamino)-pyrazolo[1,5-a]pyridine-5-carboxylicacid (2-hydroxyethoxy)amide (73a)

Compound 73a, where W=Br, Y=Cl, and X═H, can be prepared as shown inFIG. 12.

Example 32

Synthesis of6-(4-bromo-2-chlorophenylamino)-7-fluoropyrazolo[1,5-a]pyridine-5-carboxylicacid (2-hydroxyethoxy)-amide (73b)

Compound 73b, where W=Br, Y=Cl, and X═F, can be prepared as shown inFIG. 12.

Example 33

Synthesis of phosphoric acidmono-(2-{[7-(4-bromo-2-chlorophenylamino)-8-chloro-imidazo[1,2-a]pyridine-6-carbonyl]-aminooxy}-ethyl)ester (74)

7-(4-bromo-2-chlorophenylamino)-8-chloroimidazo[1,2-a]pyridine-6-carboxylicacid (2-hydroxyethoxy)-amide (33a) (100 mg, 0.234 mmol), tetrazole (23mg, 0.327 mmol) and di-tert-butyl diisopropylphosphoramidite (0.096 mL,0.304 mmol) were dissolved/suspended in 30 mL of anhydrous DMF under anatmosphere of dry N₂. The reaction mixture was stirred for about 3hours, after which time the reaction was cooled to −78° C. andtert-butyl hydrogen peroxide (0.100 mL of 70% solution in water) wasadded. The cooling bath was then taken away and the reaction was slowlywarmed up to room temperature and reacted over night. The reactionmixture was then partitioned between a solution of ethyl ether/ethylacetate (5:1) and saturated aqueous NaHCO₃. The organic layer was savedand successively washed with 10% aqueous sodium sulfite, 3 times withwater and finally with brine. The resulting organic layer was dried overMgSO₄, filtered and concentrated under vacuum. The residue was dissolvedin 3 mL of a solution of TFA/DCM (2:1) under an atmosphere of dry N₂.The reaction was stirred at room temperature for about 2 hours afterwhich time it was concentrated under vacuum and the resulting residuewas stirred in methanol for about 1 hour. The off-white solid wascollected via suction filtration, washed with methanol followed by ethylether and then air-dried to give the desired compound (74).

Example 347-(4-Bromo-2-chlorophenylamino)-8-methylimidazo[1,2-a]pyridine-6-carboxylicacid

4,6-Dichloro-5-methylnicotinic acid (J. Heterocyclic Chemistry 1999, 36,953-957) was converted to7-(4-bromo-2-chloro-phenylamino)-8-methylimidazo[1,2-a]pyridine-6-carboxylicacid according to the steps described in the alternate synthesis of StepD, Example 9. It was determined that addition of sodium azide to the4-(4-bromo-2-chloro-phenylamino)-6-chloro-5-methylnicotinic acid methylester intermediate required heating to 50° C., which results in aseparable mixture of the desired methyl ester,6-azido-4-(4-bromo-2-chlorophenylamino)-5-methylnicotinic acid methylester, and the corresponding carboxylic acid. MS ESI (+) m/z 380, 382(M+, Cl, Br pattern) detected. ¹H NMR (400 MHz, DMSO-d₆) δ 9.46 (s, 1H),9.40 (br s, 1H), 8.25 (d, 1H), 8.12 (d, 1H), 7.79 (m, 1H), 7.42 (dd,1H), 6.80 (d, 1H), 2.07 (s, 3H).

The following compound was prepared as described in the above exampleusing 4-bromo-2-fluorophenylamine in the first step:

7-(4-Bromo-2-fluorophenylamino)-8-methylimidazo[1,2-a]pyridine-6-carboxylicacid

MS ESI (+) m/z 364, 366 (M+, Cl, Br pattern) detected. ¹H NMR (400 MHz,DMSO-d₆) δ 9.40 (s, 1H), 9.26 (br s, 1H), 8.22 (d, 1H), 8.10 (d, 1H),7.61 (dd, 1H), 7.29 (dd, 1H), 6.87 (t, 1H), 2.14 (s, 3H). ¹⁹F (376 MHz,DMSO-d₆)−125.7 (s).

Example 35 Preparation of[6-(5-Amino-[1,3,4]oxadiazol-2-yl)-8-chloro-imidazo[1,2-a]pyridin-7-yl]-(4-bromo-2-fluorophenyl)-amine

Step A: Preparation of7-(4-bromo-2-fluorophenylamino)-8-chloroimidazo[1,2-a]pyridine-6-carboxylicacid hydrazide.7-(4-Bromo-2-fluoro-phenylamino)-8-chloro-imidazo[1,2-a]pyridine-6-carboxylicacid was converted to the hydrazide according to the coupling conditionsdescribed in Step A of Example 3. Alternatively, the hydrazide can beprepared directly from7-(4-Bromo-2-fluorophenylamino)-8-chloroimidazo[1,2-a]pyridine-6-carboxylicacid methyl ester by refluxing with hydrazine in ethanol. MS ESI (+) m/z398, 400 (M+, Cl, Br pattern) detected.

Step B: Preparation of[6-(5-amino-[1,3,4]oxadiazol-2-yl)-8-chloroimidazo[1,2-a]pyridin-7-yl]-(4-bromo-2-fluorophenyl)-amine.7-(4-Bromo-2-fluoro-phenylamino)-8-chloro-imidazo[1,2-a]pyridine-6-carboxylicacid hydrazide (100 mg, 0.25 mmol) was suspended in dioxane (2 mL).Cyanogen bromide (27 mg, 0.253 mmol) was added, followed by a solutionof sodium bicarbonate (21 mg, 0.25 mmol) in H₂O (1.2 mL). The reactionmixture was stirred at room temperature for 16 hours. The reactionmixture was diluted with ethyl acetate and washed with water andsaturated aqueous NaCl, dried over Na₂SO₄ and concentrated to yield thedesired product (97 mg, 91%) as a white solid. MS ESI (+) m/z 423, 425(M+, Cl, Br pattern) detected. ¹H NMR (400 MHz, CD₃OD) δ 8.98 (s, 1H),7.95 (s, 1H), 7.61 (s, 1H), 7.33 (d, 1H), 7.17 (d, 1H), 6.78 (t, 1H).¹⁹F (376 MHz, CD₃OD)−128.6 (t).

Example 36 Preparation of5-[7-(4-Bromo-2-fluorophenylamino)-8-chloroimidazo[1,2-a]pyridin-6-yl]-[1,3,4]oxadiazole-2-thiol

7-(4-Bromo-2-fluoro-phenylamino)-8-chloro-imidazo[1,2-a]pyridine-6-carboxylicacid hydrazide (50 mg, 0.13 mmol) was suspended in ethanol (2.5 mL) andcooled to 0° C. Carbon disulfide (22 mg, 0.29 mmol) was added, followedby powdered potassium hydroxide (7 mg, 0.13 mmol). The reaction mixturewas stirred under N₂ for 1 hour at 0° C. and then for 30 minutes at roomtemperature. The reaction mixture was then brought to reflux and stirredunder N₂ for 5 days. The reaction mixture was diluted with water andacidified to pH 1-2 with aqueous 1 M HCl. This mixture was thenextracted with ethyl acetate (2×). The combined organic layers werewashed with saturated aqueous NaHCO₃, saturated aqueous NaCl, dried overNa₂SO₄ and concentrated. Purification of the crude product was achievedby flash column chromatography (gradient of dichloromethane to 15:1dichloromethane/methanol) and then trituration with diethyl ether anddichloromethane to yield the desired product (17 mg, 31%) as a yellowsolid. MS ESI (+) m/z 440, 442 (M+, Cl, Br pattern) detected. ¹H NMR(400 MHz, DMSO-d₆) δ 9.29 (s, 1H), 8.16 (s, 1H), 8.08 (br s, 1H), 7.73(s, 1H), 7.49 (d, 1H), 7.15 (d, 1H), 6.59 (t, 1H). ¹⁹F (376 MHz,DMSO-d₆)−128.7 (s).

Example 37 Preparation of5-[7-(4-bromo-2-fluorophenylamino)-8-fluoroimidazo[1,2-a]pyridin-6-yl]-3H-[1,3,4]oxadiazol-2-one

7-(4-bromo-2-fluorophenylamino)-8-fluoroimidazo[1,2-a]pyridine-6-carboxylicacid hydrazide (373 mg, 0.98 mmol), which was prepared according to StepA, Example 35, was dissolved into dimethylformamide (5 mL).Carbonyldiimidazole (166 mg, 1.02 mmol) was added as a solid. Thereaction mixture was stirred for 1 hour at room temperature. It was thendiluted with ethyl acetate and washed with water. The aqueous layer wasback-extracted with ethyl acetate (3×). The combined organic layers werewashed with saturated aqueous NaCl, dried over Na₂SO₄ and concentrated.Purification of the crude product was achieved by trituration with ethylacetate and diethyl ether. The resulting solid was filtered, washed withdiethyl ether, collected and dried under vacuum. The filtrate wasconcentrated and the trituration procedure was repeated. The solids werecombined to yield the desired product (334 mg, 84%) as a yellow solid.MS ESI (+) m/z 408, 410 (M+, Br pattern) detected. ¹H NMR (400 MHz,DMSO-d₆) δ 12.69 (br s, 1H), 9.05 (s, 1H), 8.11 (d, 1H), 7.89 (s, 1H),7.67 (s, 1H), 7.51 (dd, 1H), 7.20 (d, 1H), 6.77 (m, 1H). ¹⁹F (376 MHz,DMSO-d₆)−128.9 (t, 1F), −139.5 (s, 1F).

Example 38 Preparation of[6-(5-Aminomethyl-[1,3,4]oxadiazol-2-yl)-8-chloroimidazo[1,2-a]pyridin-7-yl]-(4-bromo-2-fluorophenyl)-amine

Step A: Preparation of7-(4-bromo-2-fluorophenylamino)-8-chloroimidazo[1,2-a]pyridine-6-carboxylicacid N′-(2-chloroacetyl)-hydrazide.7-(4-Bromo-2-fluorophenylamino)-8-chloroimidazo[1,2-a]pyridine-6-carboxylicacid hydrazide (100 mg, 0.25 mmol) of Step A, Example 35 was suspendedin dichloromethane (2 mL) and 4-Me morpholine (0.040 mL, 0.36 mmol) wasadded. The mixture was cooled to 0° C. and then chloroacetyl chloride(0.029 mL, 0.36 mmol) was added dropwise. The reaction mixture waswarmed to room temperature and stirred for 1 hour under N₂. Rinsedreaction mixture into a separatory funnel with a small amount oftetrahydrofuran and methanol and then diluted with ethyl acetate. Theorganic layer was washed with water and saturated aqueous NaCl, driedover Na₂SO₄ and concentrated. Purification of the crude product wasachieved by flash column chromatography (20:1 dichloromethane/methanol)to yield the desired product (64 mg, 54%) as a yellow solid. MS ESI (+)m/z 474, 476 (M+, Cl, Br pattern) detected.

Step B: Preparation of(4-bromo-2-fluorophenyl)-[8-chloro-6-(5-chloromethyl-[1,3,4]oxadiazol-2-yl)-imidazo[1,2-a]pyridin-7-yl]-amine.7-(4-Bromo-2-fluorophenylamino)-8-chloroimidazo[1,2-a]pyridine-6-carboxylicacid N′-(2-chloro-acetyl)-hydrazide (63 mg, 0.13 mmol) was suspended inPOCl₃ (1 mL). The reaction mixture was heated to 100° C. for 8 hours,during which time it became a bright red solution. The reaction mixturewas cooled to room temperature and the solvent evaporated under reducedpressure. The residue was dissolved in ethyl acetate and washed withsaturated aqueous NaHCO₃, saturated aqueous NaCl, dried over Na₂SO₄ andconcentrated. The crude product (38 mg, 62%) was used without furtherpurification in next step. MS ESI (+) m/z 456, 458 (M+, Cl, Br pattern)detected.

Step C: Preparation of[6-(5-aminomethyl-[1,3,4]oxadiazol-2-yl)-8-chloroimidazo[1,2-a]pyridin-7-yl]-(4-bromo-2-fluorophenyl)-amine.(4-Bromo-2-fluoro-phenyl)-[8-chloro-6-(5-chloromethyl-[1,3,4]oxadiazol-2-yl)-imidazo[1,2-a]pyridin-7-yl]-amine(38 mg, 0.083 mmol) was dissolved in tetrahydrofuran (1 mL). Potassiumiodide (14 mg, 0.083 mmol) was added and then ammonia (7 M solution inmethanol, 0.30 mL, 2.08 mmol). The reaction mixture was stirred at roomtemperature for 16 hours. The solvent was removed under reduced pressureand the crude product was purified by flash column chromatography(gradient of 20:1 dichloromethane/methanol to 5:1) to yield the desiredproduct (26 mg, 71%) as a yellow solid. MS ESI (+) m/z 437, 439 (M+, Cl,Br pattern) detected. ¹H NMR (400 MHz, DMSO-d₆) δ 9.35 (s, 1H), 8.33 (s,1H), 8.17 (d, 1H), 7.70 (d, 1H), 7.51 (dd, 1H), 7.16 (d, 1H), 6.65 (t,1H), 3.94 (s, 2H). ¹⁹F (376 MHz, DMSO-d₆)−128.3 (t).

Example 392-{5-[7-(4-Bromo-2-fluorophenylamino)-8-fluoroimidazo[1,2-a]pyridin-6-yl]-[1,3,4]oxadiazol-2-ylamino}-ethanol

5-[7-(4-Bromo-2-fluoro-phenylamino)-8-fluoroimidazo[1,2-a]pyridin-6-yl]-3H-[1,3,4]oxadiazol-2-oneof Example 37 was converted in two steps to the desired productfollowing the procedures described in WO 04/056789. MS ESI (+) m/z 451,453 (M+, Br pattern) detected. ¹H NMR (400 MHz, DMSO-d₆) δ 8.96 (s, 1H),8.42 (s, 1H), 8.13 (d, 1H), 7.98 (t, 1H), 7.64 (d, 1H), 7.55 (dd, 1H),7.26 (d, 1H), 6.87 (m, 1H), 4.78 (t, 1H), 3.56 (q, 2H), 3.30 (m, 2H).¹⁹F (376 MHz, DMSO-d₆)−128.3 (t, 1F), −139.6 (s, 1F).

Example 40N-{5-[7-(4-bromo-2-fluorophenylamino)-8-fluoroimidazo[1,2-a]pyridin-6-yl]-[1,3,4]oxadiazol-2-yl}-N′-methyl-ethane-1,2-diamine

5-[7-(4-Bromo-2-fluorophenylamino)-8-fluoroimidazo[1,2-a]pyridin-6-yl]-3H-[1,3,4]oxadiazol-2-oneof Example 37 was converted in three steps to the desired product,isolated as the HCl salt, following the procedures described in WO04/056789. MS ESI (+) m/z 464, 466 (M+, Br pattern) detected. ¹H NMR(400 MHz, DMSO-d₆) δ 9.26 (s, 1H), 8.95 (br s, 1H), 8.86 (br s, 2H),8.39 (t, 1H), 8.25 (s, 1H), 7.93 (s, 1H), 7.64 (dd, 1H), 7.36 (d, 1H),7.13 (m, 1H), 3.59 (q, 2H), 3.17 (m, 2H), 2.59 (t, 3H).

Example 41 Preparation of Hydroxylamines

Hydroxylamines useful for synthesizing compounds of the presentinvention may be prepared as follows:

(i). (S)—O-[2-(tert-Butyl-dimethylsilanyloxy)-propyl]-hydroxylamine and(R)—O-[2-(tert-butyl-dimethylsilanyloxy)-propyl]-hydroxylamine

Step A: Preparation of (S)-1-iodopropan-2-ol and (R)-1-Iodo-propan-2-ol.Acetic acid (12.8 mL, 224 mmol) and (S)-(−)- or (R)-(+)-propylene oxide(16.0 mL, 224 mmol) were added sequentially to a solution of lithiumiodide (15.0 g, 112 mmol) in THF (200 mL) cooled to 0° C. The resultingthick suspension was allowed to warm to room temperature and stir for 16hours. The suspension was diluted with ether, washed with water (3×),saturated NaHCO₃ (3×), brine, dried over Na₂SO₄, and concentrated toprovide the desired product as a yellow liquid (19.5 g, 94%).

Step B: Preparation of(S)-tert-butyl-(2-iodo-1-methylethoxy)-dimethylsilane and(R)-tert-Butyl-(2-iodo-1-methylethoxy)-dimethylsilane. Pyridine (9.50mL, 118 mmol) was added to a solution of (S)— or (R)-1-iodo-propan-2-ol(19.9 g, 107 mmol) and TBSCI (17.0 g, 113 mmol) in DMF (100 mL) cooledto 0° C. After stirring for two days, the solution was diluted withhexanes, washed with water (3×) and brine. The aqueous washes wereback-extracted with 1:1 hexanes/ethyl acetate. The combined organicphases were dried over Na₂SO₄. Concentration provided the desiredproduct as a yellow liquid (26.7 g, 83%).

Step C: Preparation of(S)-2-[2-(tert-Butyl-dimethylsilanyloxy)-propoxy]-isoindole-1,3-dioneand(R)-2-[2-(tert-Butyl-dimethylsilanyloxy)-propoxy]-isoindole-1,3-dione. Asolution of (S)— or(R)-tert-butyl-(2-iodo-1-methylethoxy)-dimethylsilane (22.7 g, 75.4mmol), N-hydroxyphthalimide (14.8 g, 90.5 mmol) anddiisopropylethylamine (15.8 mL, 90.5 mmol) was heated at 75° C. for 48hours. The solution was cooled to room temperature, diluted with waterand extracted with 1:1 hexanes/ethyl acetate (2×) and diethyl ether(2×). The combined organic extracts were washed with water (3×), brine(2×), dried over Na₂SO₄ and concentrated to a red liquid. The crudeproduct was purified by flash column chromatography (dichloromethane) toprovide the desired product as a yellow liquid (12.8 g, 50%).

Step D: Preparation of(S)—O-[2-(tert-Butyl-dimethylsilanyloxy)-propyl]-hydroxylamine or(R)—O-[2-(tert-Butyl-dimethylsilanyloxy)-propyl]-hydroxylamine. Methylhydrazine (2.12 mL, 39.9 mmol) was added to a solution of (S)— or(R)-2-[2-(tert-butyl-dimethylsilanyloxy)-propoxy]-isoindole-1,3-dione(12.8 g, 38.0 mmol) in dichloromethane (100 mL) and the resultingsuspension was stirred for 16 hours. The suspension was filtered toremove solids and the filtrate was concentrated to a yellow liquid.Flash column chromatography (2:1 hexanes/ethyl acetate) provided thedesired product as a light yellow liquid (6.37 g, 82%). MS APCI (+) m/z206 (M+1) detected. ¹H NMR (400 MHz, CDCl₃) δ 5.44 (br s, 2H), 4.03 (m,1H), 3.58 (m, 1H), 3.51 (m, 1H), 1.12 (d, 3H), 0.90 (s, 9H), 0.08 (s,6H).

(ii). The following hydroxylamines were prepared similarly starting withthe appropriate terminal epoxide. The isoindol-1,3-diones intermediatesand the final hydroxylamines were purified by flash columnchromatography.

(S)-O-[2-(tert-butyl-dimethyl-silanyloxy)-butyl]-hydroxylamine and(R)-O-[2-(tert-butyl-dimethyl-silanyloxy)-butyl]-hydroxylamine

(S)—O-[2-(tert-Butyl-dimethyl-silanyloxy)-butyl]-hydroxylamine and(R)—O-[2-(tert-butyl-dimethyl-silanyloxy)-butyl]-hydroxylamine wereprepared from the homochiral terminal epoxides (S)— and(R)-1,2-epoxybutane respectively, which were obtained by kineticresolution of 1,2-epoxybutane as described within J. Am. Chem. Soc.,2002, 124:1307. MS APCI (+) m/z 220 (M+1) detected. ¹H NMR (400 MHz,CDCl₃) δ 5.41 (br s, 2H), 3.79 (m, 1H), 3.60 (m, 2H), 1.54 (m, 1H), 1.44(m, 1H), 0.90 (s, 9H), 0.08 (s, 3H), 0.07 (s, 3H).

O-[2-(tert-Butyl-dimethylsilanyloxy)-3-methylbutyl]-hydroxylamine

MS APCI (+) m/z 234 (M+1) detected. ¹H NMR (400 MHz, CDCl₃) δ 5.38 (brs, 2H), 3.64 (m, 3H), 1.75 (m, 1H), 0.90 (m, 15H), 0.08 (s, 3H), 0.05(s, 3H).

(iii). 1-Aminooxy-3,3-dimethylbutan-2-ol

Step A: Preparation of2-(2-hydroxy-3,3-dimethylbutoxy)-isoindole-1,3-dione. To a solution of3,3-dimethyl-1,2-epoxybutane (5.0 mL, 41.0 mmol) in DMF (100 mL) wasadded N-hydroxyphthalimide (8.03 g, 49.2 mmol) and triethylamine (6.90mL, 49.2 mmol). The solution was heated at 75° C. for two days. Thesolution was cooled to room temperature, diluted with ethyl acetate andwashed with water (2×), saturated potassium carbonate (3×), brine (2×),dried over Na₂SO₄ and concentrated to an orange solid. Purificationusing flash column chromatography (dichloromethane) provided the desiredproduct as a white solid (1.50 g, 14%).

Step B: Preparation of 1-aminooxy-3,3-dimethylbutan-2-ol. To a solutionof 2-(2-hydroxy-3,3-dimethylbutoxy)-isoindole-1,3-dione (1.47 g, 5.60mmol) in dichloromethane (20 mL) cooled to 0° C. was addedmethylhydrazine (0.31 mL, 5.90 mmol). The white suspension was allowedto stir for 16 hours at room temperature. Diethyl ether (50 mL) wasadded and the solids were removed by filtration. The filtrate wasconcentrated, diluted with diethyl ether and the solids were removed byfiltration. This procedure was repeated twice more and the finalfiltrate was concentrated to provide the desired product as a yellowliquid (0.643 g, 86%). ¹H NMR (400 MHz, CDCl₃) δ 4.87 (br s, 2H), 3.85(q, 1H), 3.58 (q, 2H), 0.93 (s, 9H).

(iv). (2-Aminooxy-ethyl)-carbamic acid tert-butyl ester

Step A: Preparation of methanesulfonic acid2-tert-butoxycarbonylamino-ethyl ester. Methanesulfonyl chloride (0.60mL, 7.76 mmol) was added to a solution of (2-hydroxy-ethyl)-carbamicacid tert-butyl ester (1.04 g, 6.46 mmol) and triethylamine (1.35 mL,9.70 mmol) in dichloromethane (35 mL) cooled to 0° C. The solution wasstirred for one hour, after which time it was diluted with ethylacetate, washed with saturated NaHCO₃ (2×), brine, dried over Na₂SO₄ andconcentrated to a thick colorless liquid (1.37 g, 89%).

Step B: Preparation of[2-(1,3-dioxo-1,3-dihydroisoindol-2-yloxy)-ethyl]-carbamic acidtert-butyl ester. Added N-hydroxyphthalimide (1.12 g, 6.87 mmol) andtriethylamine (0.96 mL, 6.87 mmol) to a solution of methanesulfonic acid2-tert-butoxycarbonylaminoethyl ester (1.37 g, 5.73 mmol) in DMF (20mL). The solution was heated to 50° C. for 16 hours after which time itwas cooled to room temperature. The solution was diluted with ethylacetate, washed with water (2×), saturated K₂CO₃, dried over Na₂SO₄ andconcentrated to an orange solid (747 mg, 43%) which was taken on withoutpurification.

Step C: Preparation of (2-aminooxyethyl)-carbamic acid tert-butyl ester.The synthesis of the title compound was carried out according to Step Dof Example 41 (i) using[2-(1,3-dioxo-1,3-dihydroisoindol-2-yloxy)-ethyl]-carbamic acidtert-butyl ester as the starting material to provide 255 mg (71%) of thedesired compound as a yellow liquid. ¹H NMR (400 MHz, CDCl₃) δ 5.50 (brs, 2H), 5.02 (br s, 1H), 3.71 (t, 2H), 3.36 (q, 2H), 1.45 (s, 9H).

The following hydroxylamine was prepared similarly using(3-hydroxy-propyl)-carbamic acid tert-butyl ester as the startingmaterial.

(3-Aminooxypropyl)-carbamic acid tert-butyl ester

¹H NMR (400 MHz, CDCl₃) δ 5.41 (br s, 2H), 4.76 (br s, 1H), 3.73 (t,2H), 3.21 (q, 2H), 1.78 (m, 2H), 1.44 (s, 9H).

(v). (3-Aminooxy-2,2-dimethylpropyl)-carbamic acid tert-butyl ester

Step A: Preparation of (3-hydroxy-2,2-dimethylpropyl)-carbamic acidtert-butyl ester. Boc-anhydride (13.07 g, 59.9 mmol) in THF (10 mL) wasadded dropwise to a solution of 3-amino-2,2-dimethylpropan-1-ol (5.15 g,49.9 mmol) and NaOH (2.40 g, 59.9 mmol) dissolved into 1:1 THF/water (50mL). The solution was stirred at room temperature for 72 hours. Thesolution was concentrated under reduced pressure to about one half ofthe reaction volume. The remaining solution was acidified to pH 6 andwas then extracted with ethyl acetate (2×). The organic extracts werewashed with water, brine, dried over Na₂SO₄ and concentrated to providethe desired product (10.2 g, quantitative) as a white solid.

Step B: Preparation of[3-(1,3-dioxo-1.3-dihydroisoindol-2-yloxy)-2,2-dimethylpropyl]-carbamicacid tert-butyl ester. Diethylazodicarboxylate (8.26 mL, 52.4 mmol) wasadded to a solution of (3-hydroxy-2,2-dimethyl-propyl)-carbamic acidtert-butyl ester (10.2 g, 49.9 mmol), N-hydroxyphthalimide (8.15 g, 49.9mmol) and triphenylphosphine (13.1 g, 49.9 mmol) in THF (200 mL). Thesolution was stirred at room temperature for 16 hours after which timeit was diluted with dichloromethane and passed through a plug of silicagel, eluting with dichloromethane. The product containing fractions wereconcentrated to a yellow liquid, which was further purified by flashcolumn chromatography (dichloromethane to 4:1 dichloromethane/ethylacetate) to provide pure desired product (1.98 g, 11%) as a waxy whitesolid.

Step C: Preparation of (3-aminooxy-2,2-dimethylpropyl)-carbamic acidtert-butyl ester. The synthesis of the title compound was carried outaccording to STEP D of the Example 41 using[3-(1,3-dioxo-1,3-dihydroisoindol-2-yloxy)-2,2-dimethylpropyl]-carbamicacid tert-butyl ester as the starting material to provide 998 mg (80%)desired product as a pale yellow liquid. ¹H NMR (400 MHz, CDCl₃) δ 5.45(br s, 2H), 4.94 (br s, 1H), 3.44 (s, 2H), 3.03 (br d, 2H), 1.45 (s,9H), 0.88 (s, 6H).

(vi). (3-Aminooxy-1-methylpropyl)-carbamic acid tert-butyl ester

Step A: Preparation of 3-aminobutan-1-ol. Lithium aluminum hydride (1.0M in THF, 43.8 mL, 43.8 mmol) was added dropwise over one hour to asuspension of 3-aminobutyric acid (2.26 g, 21.9 mmol) in THF (100 mL)cooled to 0° C. The solution was then refluxed for 16 hours after whichtime it was cooled to 0° C. and quenched by the careful sequentialaddition of water (2 mL), 15% aqueous NaOH (2 mL) and water (2 mL). Themixture was stirred for 15 minutes and was filtered through CELITE,washing the filter pad with THF. Concentration of the filtrated providedthe desired product (1.43 g, 73%) as a clear oil.

Step B: Preparation of (3-aminooxy-1-methylpropyl)-carbamic acidtert-butyl ester. The synthesis of the title compound was carried outaccording to Steps A, B and C of Example 41(iii) above using3-aminobutan-1-ol as the starting material to provide 998 mg (80%)desired product as a pale yellow liquid. ¹H NMR (400 MHz, CDCl₃) δ 5.39(br s, 2H), 4.52 (br s, 1H), 3.72 (m, 3H), 1.70 (m, 2H), 1.43 (2, 9H),1.14 (d, 3H).

(vii). The following hydroxylamines were prepared as described in WO02/06213: O-(2-vinyloxyethyl)-hydroxylamine;O-(2-methoxyethyl)-hydroxylamine; 2-aminooxypropan-1-ol;3-aminooxypropan-1-ol; 1-aminooxy-2-methylpropan-2-ol;1-aminooxy-3-methoxypropan-2-ol; 3-aminooxy-1,1,1-trifluoropropan-2-ol;2-aminooxy-2methylpropan-1-ol; (2-aminooxyethyl)-methylcarbamic acidtert-butyl ester;(R)—O-2,2-dimethyl-[1,3]dioxolan-4-ylmethyl)-hydroxylamine;(S)—O-2,2-dimethyl-[1,3]dioxolan-4-ylmethyl)-hydroxylamine.

(viii). The isoindole-1,3-dione intermediates of the followinghydroxylamines are prepared from the appropriate alkyl halide andN-hydroxyphthalimide by the procedure described within J. HeterocyclicChemistry 2000, 37, 827-830: O-propyl-hydroxylamine;O-isopropylhydroxylamine; O-cyclopropylmethylhydroxylamine. Theisoindole-1,3-diones are deprotected by the procedure described above.

(ix). The following hydroxylamines were obtained from commercialsources: methoxylamine hydrochloride; O-ethylhydroxylaminehydrochloride; O-(tert-butyl)amine hydrochloride; O-allylaminehydrochloride.

Additional compounds suitable for use in the methods of the presentinvention are shown in FIGS. 13A-13G.

Biological Examples

Drugs

The preparation of AR-14[7-(4-bromo-2-chlorophenylamino)-8-chloroimidazo[1,2-a]pyridine-6-carboxylicacid (2-hydroxyethoxy)-amide]and AR-13[7-(4-bromo-2-fluorophenylamino)-8-chloroimidazo[1,2-a]pyridine-6-carboxylicacid (2-hydroxyethoxy)-amide] is described in the above Examples.

Example 42 Effect of AR-14 on Collagen-Induced Arthritis (CIA)

Type II collagen-induced arthritis (CIA) in rats is an experimentalmodel of arthritis that has a number of pathologic, immunologic, andgenetic features in common with rheumatoid arthritis.

Animals

Female Lewis rats (7-9 weeks of age; 8 per group for arthritis, 4 pergroup for normal control) were acclimated for 7 days after arrival.

Experimental Protocol

Acclimated animals were anesthetized with Isoflurane. The disease wasinduced by immunization of the rats with type II collagen (equal mixtureof 4 mg/mL bovine type II collagen in 0.01 N acetic acid and Freund'sincomplete adjuvant) by injection (day 0). On day 6, the rats wereanesthetized again for the second collagen injection. Each animalreceived 300 μL of the collagen mixture on days 0 and 6, which wasspread over 3 sites on the back.

Caliper measurements of normal (pre-disease) right and left ankle jointswere taken on day 9 post-collagen injection. On days 10-11, onset ofarthritis occurred and rats were randomized into treatment groups.Randomization into each group was done after ankle joint swelling wasestablished in at least one hind paw.

Once enrolled in the treatment groups, treatment with once daily oraldosing of AR-14 (1, 3, 10, 30 or 60 mg/kg) or indomethacin (1 mg/kg) wasinitiated. Animals receiving vehicle or compound were enrolled and dailydosing was initiated using a volume of 5 mL/kg for oral solutions.ENBREL (etanercept) treatment was on days 11 and 15 (10 mg/kg) bysubcutaneous administration. Rats were weighed on days 11-18 and calipermeasurements of ankles were taken every day. Final body weights weretaken on day 19. On day 19, animals were euthanized, both hind paws andknees were removed, hind paws were weighed and then (with knees) placedin formalin.

Following 1-2 days in fixative and then 4-5 days in decalcifier, theankle joints and knees were processed, embedded, sectioned and stainedwith toluidine blue.

Scoring of Joints

A clinical scoring index was used to assess disease progression.Collagen arthritic ankles and knees were given scores of 0-5 forinflammation, pannus formation and bone resorption according to thecriteria in Tables 1-7.

TABLE 1 Knee and Ankle Inflammation Score Evaluation 0 Normal 1 Minimalinfiltration of inflammatory cells in periarticular tissue 2 Mildinfiltration 3 Moderate infiltration with moderate edema 4 Markedinfiltration with marked edema 5 Severe infiltration with severe edema

TABLE 2 Ankle Pannus (Emphasis on tibiotarsal joint) Score Evaluation 0Normal 1 Minimal infiltration of pannus in cartilage and subchondralbone 2 Mild infiltration (<¼ of tibia at edges) 3 Moderate infiltration(¼ to ⅓ of tibia affected, smaller tarsals affected) 4 Markedinfiltration (½-¾ of tibia affected, destruction of smaller tarsals) 5Severe infiltration (>¾ of tibia affected, severe distortion of overallarchitecture)

TABLE 3 Knee Pannus Score Evaluation 0 Normal 1 Minimal infiltration ofpannus in cartilage and subchondral bone 2 Mild infiltration (extendsover up to ¼ of surface or subchondral area of tibia or femur) 3Moderate infiltration (extends over >¼ but <½ of surface or subchondralarea of tibia or femur) 4 Marked infiltration (extends over ½ to ¾ oftibial or femoral surface) 5 Severe infiltration (covers >¾ of surface)

TABLE 4 Cartilage Damage (Ankle) Score Evaluation 0 Normal 1 Minimal tomild loss of toluidine blue staining with no obvious chondrocyte loss orcollagen disruption 2 Mild loss of toluidine blue staining with focalmild (superficial) chondrocyte loss and/or collagen disruption and fulldestruction of tibia <¼ of surface, mild changes in smaller tarsals 3Moderate loss of toluidine blue staining with multifocal moderate (depthto middle zone) chondrocyte loss and/or collagen disruption, ¼ to ⅓ oftibia affected by full thickness destruction, smaller tarsals affectedto ½-¾ depth 4 Marked loss of toluidine blue staining with multifocalmarked (depth to deep zone) chondrocyte loss and/or collagen disruption,½-¾ of tibia with full thickness destruction, destruction of smallertarsals 5 Severe diffuse loss of toluidine blue staining with multifocalsevere (depth to tide mark) chondrocyte loss and/or collagen disruption

TABLE 5 Cartilage Damage (Knee, emphasis on femoral condyles) ScoreEvaluation 0 Normal 1 Minimal to mild loss of toluidine blue stainingwith no obvious chondrocyte loss or collagen disruption 2 Mild loss oftoluidine blue staining with focal mild (superficial) chondrocyte lossand/or collagen disruption 3 Moderate loss of toluidine blue stainingwith multifocal to diffuse moderate (depth to middle zone) chondrocyteloss and/or collagen disruption 4 Marked loss of toluidine blue stainingwith multifocal to diffuse marked (depth to deep zone) chondrocyte lossand/or collagen disruption 5 Severe diffuse loss of toluidine bluestaining with multifocal severe (depth to tide mark) chondrocyte lossand/or collagen disruption on both femur and tibia

TABLE 6 Bone Resorption (Ankle) Score Evaluation 0 Normal 1 Minimal,i.e., small areas of resorption, not readily apparent on lowmagnification, rare osteoclasts 2 Mild, i.e., more numerous areas ofresorption, not readily apparent on low magnification, osteoclasts morenumerous, <¼ of tibia at edges is resorbed 3 Moderate, i.e., obviousresorption of medullary trabecular and cortical bone without fullthickness defects in cortex, loss of some medullary trabeculae, lesionapparent on low magnification, osteoclasts more numerous, ¼ to ⅓ oftibia affected, smaller tarsals affected 4 Marked, i.e., full thicknessdefects in cortical bone, often with distortion of profile of remainingcortical surface, marked loss of medullary bone, numerous osteoclasts,½-¾ of tibia affected, destruction of smaller tarsals 5 Severe, i.e.,full thickness defects in cortical bone, often with distortion ofprofile of remaining cortical surface, marked loss of medullary bone,numerous osteoclasts, >¾ of tibia affected, severe distortion of overallarchitecture

TABLE 7 Bone Resorption (Knee) Score Evaluation 0 Normal 1 Minimal,i.e., small areas of resorption, not readily apparent on lowmagnification, rare osteoclasts 2 Mild, i.e., more numerous areas ofresorption, definite loss of subchondral bone involving ¼ of tibial orfemoral surface (medial or lateral) 3 Moderate, i.e., obvious resorptionof subchondral bone involving >¼ but <½ of tibial or femoral surface(medial or lateral) 4 Marked, i.e., obvious resorption of subchondralbone involving ≧½ but <¾ of tibial or femoral surface (medial orlateral) 5 Severe, i.e., distortion of entire joint due to destructioninvolving >¾ of tibial or femoral surface (medial or lateral)

Statistical Analysis

Clinical data for ankle joint diameter were analyzed by determining thearea under the dosing curve (AUC). For calculation of AUC, the dailymeasurements of ankle joints (using a caliper) for each rat were enteredinto Microsoft Excel and the area between the treatment days after theonset of disease to the termination day was computed. Means for eachgroup were determined and percent inhibition from arthritis controls wascalculated by comparing values for treated and normal animals. Data wereanalyzed by the Mann Whitney or Student's t test. Paw weights andhistologic parameters (mean±SE) for each group were analyzed fordifferences using the Mann Whitney or Student's t test. In both cases,significance was set at p≦0.05.

Percent inhibition of paw weight and AUC was calculated using thefollowing formula:% Inhibition=A−(B/A)×100

wherein A=(Mean Disease Control−Mean Normal) and B=(Mean Treated−MeanNormal).

Results

AR-14 was well tolerated at all doses administered. FIG. 1 shows theeffects of AR-14 on paw diameter over 7 days. AR-14 significantlyinhibited ankle swelling compared to vehicle control at all dosestested. AR-14 stopped the progression of established disease at 10 mg/kgand caused improvement to normal at 60 mg/kg. The activity of AR-14 at10 mg/kg was equivalent to indomethacin (Arth+Indo) and better than thatof ENBREL (etanercept) (Arth+Enbrel) in this model.

FIG. 2 shows the effects of AR-14 versus indomethacin (Indo) and ENBREL(etanercept) on body weight after 7 days. AR-14 significantly inhibitedbody weight loss at all doses tested with near normal body weightsobserved at doses of 30 and 60 mg/kg. The ED₅₀ was 4.57 mg/kg.

FIG. 3 shows the effects of AR-14 versus indomethacin (Indo) and ENBREL(etanercept) on ankle histopathology after 7 days. AR-14 significantlyinhibited pannus formation, cartilage damage and bone resorption at 1and 3 mg/kg. Near normal joints with minimal inflammation at 10 mg/kgwas observed, which is significantly better than indomethacin orENBREL(etanercept) at this dose.

In summary, AR-14 significantly inhibited the progression of establishedarthritis at doses as low as 10 mg/kg and significantly inhibited jointdestruction at a dose of 1 mg/kg.

Example 43 Effect of AR-14 on Adjuvant-Induced Arthritis (AIA)

Animals

Male Lewis rats (7-9 weeks of age; 8 per group for arthritis, 4 pergroup for normal control) were acclimated for 7 days after arrival.

Experimental Protocol

Acclimated animals were randomized into groups by body weight and thenanesthetized with Isoflurane and given lipoidal amine (LA) in Freund'scomplete adjuvant (100 μL injection of 7 mg LA, intradermaladministration at the base of the tail, Day 0).

Caliper measurements of normal (pre-disease) right and left ankle jointswere taken on day 7 post-adjuvant injection. On day 8, dosing began forvehicle and AR-14 treatment groups. Methotrexate treatment (0.075 mg/kg)began on day 0 prior to adjuvant injection.

Once enrolled in the treatment group, treatment with once daily oraldoses of AR-14 (1, 3, 10 or 30 mg/kg) was initiated. AR-14 was welltolerated at all doses administered. Animals receiving vehicle orcompound were enrolled and daily dosing was initiated using a volume of5 mL/kg for oral solutions on days 8-14. Rats were weighed on days 8-14and caliper measurements of ankles were taken every day. Final bodyweights and caliper measurements of ankles and paws were taken on day15. On day 15, animals were euthanized, both hind paws were removed, andthe hind paws weighed and then placed in formalin for histopathologicanalyses. Spleens were also removed, weighed and placed in formalin forhistopathologic analyses.

Results

FIG. 4 shows the effects of AR-14 versus methotrexate (MTX) on pawdiameter over 15 days. AR-14 significantly inhibited ankle swellingcompared to vehicle control at all doses tested. AR-14 stopped theprogression of established disease and causes improvement to normal at30 mg/kg. The activity of AR-14 at 30 mg/kg was equivalent tomethotrexate in this model.

FIG. 5 shows the effects of AR-14 versus methotrexate (MTX) on bodyweight after 15 days. AR-14 significantly inhibited body weight loss at30 mg/kg.

FIG. 6 shows the effects of AR-14 versus methotrexate (MTX) on pawweight after 15 days. AR-14 significantly inhibited paw weight gain(edema/inflammation) at all doses compared to vehicle control. Nearnormal joints with minimal inflammation were observed at 30 mg/kg(comparable to methotrexate).

FIG. 7 shows the effects of AR-14 versus methotrexate (MTX) on relativespleen weight after 15 days. AR-14 significantly inhibited splenomegalyat 3, 10 and 30 mg/kg. At the latter two doses, this inhibition iscomparable to that seen with methotrexate.

Example 44 Effect of AR-14 on Indomethacin-Induced Inflammatory BowelDisease (IBD)

This study evaluated AR-14 (PO, QD) in a model of Crohn's disease(indomethacin induced intestinal injury) in rats to determine ifbeneficial effects could be detected in this IBD model.

Experimental Protocol

Rats received two subcutaneous administrations of indomethacin (7.5mg/kg, in 5% NaHCO₃) spaced 24 hours apart. Rats received once dailyoral doses of vehicle, AR-14 (1, 3, 10 or 30 mg/kg) or dexamethasone(0.1 mg/kg). Rats were euthanized and evaluated for effects of treatmentat day 4 post-initial indomethacin injection. Body weights were taken aswell as intestinal weights per 10 cm of area at risk.

Scoring of Intestines

A clinical scoring index (Table 8) was used to assess diseaseprogression. Intestines were scored for gross lesions and multiplesections of gut taken for histology (5 sections per rat spaced equallyalong area at risk).

TABLE 8 Gross Scoring Criteria Score Evaluation 0 Normal 1 Minimalthickening 2 Mild to moderate small intestinal/mesenteric thickening 3Moderate thickening with 1 or more definite adhesions that would be easyto separate 4 Marked thickening with numerous hard to separate adhesions5 Severe intestinal lesion resulting in death

Histologic Scoring Criteria

Five equally spaced sections from area at risk were scored for necrosisand inflammation (0,0) (1,3) etc., according to the criteria in Tables 9and 10.

TABLE 9 Necrosis Score Percent of lesion with necrosis 0 Normal 1 10% 210-25% 3 25-50% 4 50-75% 5  75-100%

TABLE 10 Inflammation Score Evaluation 0 Normal 1 Minimal in mesenteryand muscle or lesion 2 Mild in mesentery and muscle or lesion 3 Moderatein mesentery and muscle or lesion 4 Marked in lesion 5 Severe in lesion

Results

FIG. 8 shows the effects of AR-14 versus dexamethasone (Dex) on smallintestine score after 4 days. AR-14 significantly inhibited gross lesionscore at all doses. At the 10 and 30 mg/kg doses, AR-14 wassignificantly better than dexamethasone at inhibiting gross lesionscore.

FIG. 9 shows the effects of AR-14 versus dexamethasone (Dex) on smallintestine weight after 4 days. AR-14 significantly inhibited smallintestine weight gain as a result of edema/inflammation at the 3, 10 and30 mg/kg doses. At the 10 and 30 mg/kg doses, AR-14 was significantlybetter than dexamethasone at inhibiting intestinal inflammation andedema as well as small intestine necrosis and inflammation.

FIG. 10 shows the effects of AR-14 versus dexamethasone (Dex) on thehistopathology of the gut (necrosis and inflammation scored). At 30mg/kg, AR-14 was significantly more efficacious than dexamethasone inthis model.

In summary, a significant inhibition of disease (gut score and weightand microscopic changes) was seen with once daily dosing of AR-14 at 10mg/kg.

Example 45 Effect of AR-13 and AR-14 on Carrageenan Paw Edema

The rat carrageenan paw edema model of inflammation was utilized toevaluate the effectiveness of AR-13 and AR-14 in ameliorating swellingand inflammation.

Animals

Male Lewis rats (7-9 weeks of age; 10 per group) were acclimated for atleast 7 days after arrival.

Experimental Protocol

Acclimated animals were randomized into groups by body weight. Animalswere treated orally with AR-13 or AR-14 (3, 10 or 30 mg/kg) or with apositive control (indomethacin, 2 mg/kg). After 2 hours, the animalswere anesthetized with Isoflurane and given an intradermal injection of1.2% carrageenan into the right hind footpad. Four hours after thecarrageenan injection, the animals were sacrificed with carbon dioxideinhalation and both paws were removed at the medial malleolus andweighed. The difference in weight between the right (carrageenan) andleft (normal) paws was calculated.

Results

FIG. 11 shows the effects of AR-13 and AR-14 versus indomethacin on pawweight. Treatment with AR-13 or AR-14 significantly inhibited paw weightat 3, 10 and 30 mg/kg. The ED₅₀ for AR-14 was about 4 mg/kg, while theED₅₀ for AR-13 was about 10 mg/kg. The activity seen with both of thesecompounds was superior to indomethacin.

The foregoing description is considered as illustrative only of theprinciples of the invention. Further, since numerous modifications andchanges will be readily apparent to those skilled in the art, it is notdesired to limit the invention to the exact construction and processshown as described above. Accordingly, all suitable modifications andequivalents may be resorted to falling within the scope of the inventionas defined by the claims that follow.

The words “comprise,” “comprising,” “include,” “including,” and“includes” when used in this specification and in the following claimsare intended to specify the presence of stated features, integers,components, or steps, but they do not preclude the presence or additionof one or more other features, integers, components, steps, or groupsthereof.

1. A method for treating an inflammatory disease or disorder, saidmethod comprising administering to a mammal in need of such treatment aneffective amount of compound of the Formula II

or a solvate, metabolite or pharmaceutically acceptable salt thereof,wherein: R¹, R², R⁷, R⁸, R⁹, R¹⁰ and R¹¹ are independently hydrogen,halo, cyano, nitro, trifluoromethyl, difluoromethoxy, trifluoromethoxy,azido, —OR³, —C(O)R³, —C(O)OR³, NR⁴C(O)OR⁶, —OC(O)R³, —NR⁴SO₂R⁶,—SO₂NR³R⁴, —NR⁴C(O)R³, —C(O)NR³R⁴, —NR⁵C(O)NR³R⁴, —NR⁵C(NCN)NR³R⁴,—NR³R⁴, C₁-C₁₀ alkyl, C₂-C₁₀ alkenyl, C₂-C₁₀ alkynyl, C₃-C₁₀ cycloalkyl,C₃-C₁₀ cycloalkylalkyl, —S(O)_(j)(C₁-C₆ alkyl),—S(O)_(j)(CR⁴R⁵)_(m)-aryl, aryl, arylalkyl, heteroaryl, heteroarylalkyl,heterocyclyl, heterocyclylalkyl, —O(CR⁴R⁵)_(m)-aryl,—NR⁴(CR⁴R⁵)_(m)-aryl, —O(CR⁴R⁵)_(m)-heteroaryl,—NR⁴(CR⁴R⁵)_(m)-heteroaryl, —O(CR⁴R⁵)_(m)-heterocyclyl or—NR⁴(CR⁴R⁵)_(m)-heterocyclyl, wherein any of said alkyl, alkenyl,alkynyl, cycloalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl,heterocyclyl and heterocyclylalkyl portions are optionally substitutedwith one or more groups independently selected from oxo, halo, cyano,nitro, trifluoromethyl, difluoromethoxy, trifluoromethoxy, azido,—NR⁴SO₂R⁶, —SO₂NR³R⁴, —C(O)R³, —C(O)OR³, —OC(O)R³, —NR⁴C(O)OR⁶,—NR⁴C(O)R³, —C(O)NR³R⁴, —NR³R⁴, —NR⁵C(O)NR³R⁴, —NR⁵C(NCN)NR³R⁴, —OR³,aryl, heteroaryl, arylalkyl, heteroarylalkyl, heterocyclyl andheterocyclylalkyl; or R⁷ and R¹¹ together with the atoms to which theyare attached form a 4 to 10 membered saturated, unsaturated, orpartially saturated carbocyclic or heterocyclic ring, wherein any ofsaid saturated, unsaturated, partially saturated carbocyclic orheterocyclic rings are optionally substituted with one or more groupsindependently selected from halo, cyano, nitro, trifluoromethyl,difluoromethoxy, trifluoromethoxy, azido, —NR′SO₂R″″, —SO₂NR′R″,—C(O)R′, —C(O)OR′, —OC(O)R′, —NR′C(O)OR″″, —NR′C(O)R″, —C(O)NR′R″,—SO₂R″″, —NR′R″, —NR′C(O)NR″R″′, —NR′C(NCN)NR″R″′, —OR′, aryl,heteroaryl, arylalkyl, heteroarylalkyl, heterocyclyl andheterocyclylalkyl; R³ is hydrogen, trifluoromethyl, C₁-C₁₀ alkyl, C₂-C₁₀alkenyl, C₂-C₁₀ alkynyl, C₃-C₁₀ cycloalkyl, C₃-C₁₀ cycloalkylalkyl,aryl, arylalkyl, heteroaryl, heteroarylalkyl, heterocyclyl,heterocyclylalkyl, phosphate or an amino acid residue, wherein any ofsaid alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkylalkyl, aryl,arylalkyl, heteroaryl, heteroarylalkyl, heterocyclyl andheterocyclylalkyl portions are optionally substituted with one or moregroups independently selected from oxo, halo, cyano, nitro,trifluoromethyl, difluoromethoxy, trifluoromethoxy, azido, —NR′SO₂R″″,—SO₂NR′R″, —C(O)R′, —C(O)OR′, —OC(O)R′, —NR′C(O)OR″″, —NR′C(O)R″,—C(O)NR′R″, —SR′, —S(O)R″″, —SO₂R″″, —NR′R″, —NR′C(O)NR″R″′,—NR′C(NCN)NR″R″′, —OR′, aryl, heteroaryl, arylalkyl, heteroarylalkyl,heterocyclyl and heterocyclylalkyl, or R³ and R⁴ together with the atomto which they are attached form a 4 to 10 membered saturated,unsaturated, or partially saturated heterocyclic ring, wherein any ofsaid saturated, unsaturated, or partially saturated heterocyclic ringsare optionally substituted with one or more groups independentlyselected from halo, cyano, nitro, trifluoromethyl, difluoromethoxy,trifluoromethoxy, azido, —NR′SO₂R″″, —SO₂NR′R″, —C(O)R′, —C(O)OR′,—OC(O)R′, —NR′C(O)OR″″, —NR′C(O)R″, —C(O)NR′R″, —SO₂R″″, —NR′R″,—NR′C(O)NR″R″′, —NR′C(NCN)NR′R″′, —OR′, aryl, heteroaryl, arylalkyl,heteroarylalkyl, heterocyclyl and heterocyclylalkyl; R′, R″ and R″′ areindependently hydrogen, lower alkyl, lower alkenyl, aryl or arylalkyl;R″″ is lower alkyl, lower alkenyl, aryl or arylalkyl, or any two of R′,R″, R″′ and R″″ together with the atoms to which they are attached forma 4 to 10 membered saturated, unsaturated, or partially saturatedheterocyclic ring, wherein any of said alkyl, alkenyl, aryl, arylalkylsaturated, unsaturated, or partially saturated heterocyclic rings areoptionally substituted with one or more groups independently selectedfrom halo, cyano, nitro, trifluoromethyl, difluoromethoxy,trifluoromethoxy, azido, aryl, heteroaryl, arylalkyl, heteroarylalkyl,heterocyclyl, and heterocyclylalkyl; R⁴ and R⁵ are independentlyhydrogen or C₁-C₆ alkyl; R⁶ is trifluoromethyl, C₁-C₁₀ alkyl, C₃-C₁₀cycloalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, heterocyclylor heterocyclylalkyl, wherein any of said alkyl, cycloalkyl, aryl,arylalkyl, heteroaryl, heteroarylalkyl, heterocyclyl andheterocyclylalkyl portions are optionally substituted with one or moregroups independently selected from oxo, halo, cyano, nitro,trifluoromethyl, difluoromethoxy, trifluoromethoxy, azido, —NR′SO₂R″″,—SO₂NR′R″, —C(O)R′, —C(O)OR′, —OC(O)R′, —NR′C(O)OR″″, —NR′C(O)R″,—C(O)NR′R″, —SO₂R″″, —NR′R′, —NR′C(O)NR″R″′, —NR′C(NCN)NR″R″′, —OR′,aryl, heteroaryl, arylalkyl, heteroarylalkyl, heterocyclyl, andheterocyclylalkyl; W is heteroaryl, heterocyclyl, —C(O)OR³, —C(O)NR³R⁴,—C(O)NR⁴OR³, C(O)R⁴OR³, —C(O)(C₃-C₁₀ cycloalkyl), —C(O)(C₁-C₁₀ alkyl),—C(O)(aryl), C(O)(heteroaryl), —C(O)(heterocyclyl), —CONH(SO₂)CH₃ or—CR³OR³, wherein any of said heteroaryl, heterocyclyl, —C(O)OR³,—C(O)NR³R⁴, —C(O)NR⁴OR³, —C(O)R⁴OR³, —C(O)(C₃-C₁₀ cycloalkyl),—C(O)(C₁-C₁₀ alkyl), —C(O)(aryl), —C(O)(heteroaryl),—C(O)(heterocyclyl), —CONH(SO₂)CH₃ and —CR³OR³ are optionallysubstituted with one or more groups independently selected from —NR³R⁴,—OR³, —R², C₁-C₁₀ alkyl, C₂-C₁₀ alkenyl and C₂-C₁₀ alkynyl, wherein anyof said alkyl, alkenyl, and alkynyl portions are optionally substitutedwith 1 or more groups independently selected from —NR³R⁴ and —OR³; m is0, 1, 2, 3, 4 or 5; j is 0, 1 or 2; and Y is a linker.
 2. The method ofclaim 1, wherein said inflammatory disorder is rheumatoid arthritis. 3.The method of claim 2, wherein said compound is administered in anamount between about 1 and 60 mg per kilogram of body weight of saidmammal.
 4. The method of claim 3, wherein said amount is between about 1and 10 mg per kilogram of body weight of said mammal.
 5. The method ofclaim 4, wherein said amount is between about 1 and 3 mg per kilogram ofbody weight of said mammal.
 6. The method of claim 1, wherein saidcompound is administered once a day.
 7. The method of claim 1, whereinsaid compound is administered orally.
 8. The method of claim 1, whereinsaid compound is7-(4-bromo-2-chlorophenylamino)-8-chloroimidazo[1,2-a]pyridine-6-carboxylicacid (2-hydroxyethoxy)-amide.
 9. The method of claim 1, wherein saidcompound is7-(4-bromo-2-fluorophenylamino)-8-chloroimidazo[1,2-a]pyridine-6-carboxylicacid (2-hydroxyethoxy)-amide.